• 产品描述Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules. Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate. Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation.
• 产品名称Anti-GAPDH Recombinant Rabbit Monoclonal Antibody [SA30-01]
• 分子量Predicted band size: 36 kDa
• 种属反应性Human,Mouse,Rat, Zebrafish
• 验证应用WB,ICC/IF,IHC-P,FC,IP
• 抗体类型重组兔单抗
• 免疫原Recombinant protein within mouse GAPDH aa 94-333 / 333.
• 偶联Non-conjugated

• 性能

• 形态Liquid
• 浓度1 mg/mL.
• 存放说明Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
• 存储缓冲液1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
• 亚型IgG
• 纯化方式Protein A affinity purified.
• 亚细胞定位Cytoplasm, Nucleus, Membrane
• 数据链接SwissProt: P04406 Human
  SwissProt: P16858 Mouse
  SwissProt: P04797 Rat
• 其它名称
  • 38 kDa BFA-dependent ADP-ribosylation substrate antibody
  • aging associated gene 9 protein antibody
  • Aging-associated gene 9 protein antibody
  more

• 应用

  WB: 1:5,000-1:640,000
  ICC/IF: 1:50
  IHC-P: 1:50
  FC: 1:50
  IP: Use at an assay dependent concentration.

产品描述Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules. Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate. Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation.

产品描述产品描述Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules. Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate. Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation.

产品名称Anti-GAPDH Recombinant Rabbit Monoclonal Antibody [SA30-01]

产品名称产品名称Anti-GAPDH Recombinant Rabbit Monoclonal Antibody [SA30-01]

分子量Predicted band size: 36 kDa

分子量分子量Predicted band size: 36 kDa

种属反应性Human,Mouse,Rat, Zebrafish

种属反应性种属反应性Human,Mouse,Rat, Zebrafish

验证应用WB,ICC/IF,IHC-P,FC,IP

验证应用验证应用WB,ICC/IF,IHC-P,FC,IP

抗体类型重组兔单抗

抗体类型抗体类型重组兔单抗

免疫原Recombinant protein within mouse GAPDH aa 94-333 / 333.

免疫原免疫原Recombinant protein within mouse GAPDH aa 94-333 / 333.

偶联Non-conjugated

偶联偶联Non-conjugated

性能

性能

性能

形态Liquid

形态形态Liquid

浓度1 mg/mL.

浓度浓度1 mg/mL.

存放说明Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

存放说明存放说明Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

存储缓冲液1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

存储缓冲液存储缓冲液1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

亚型IgG

亚型亚型IgG

纯化方式Protein A affinity purified.

纯化方式纯化方式Protein A affinity purified.

亚细胞定位Cytoplasm, Nucleus, Membrane

亚细胞定位亚细胞定位Cytoplasm, Nucleus, Membrane

数据链接SwissProt: P04406 Human
SwissProt: P16858 Mouse
SwissProt: P04797 Rat

数据链接数据链接SwissProt: P04406 HumanSwissProt: P04406 Human
SwissProt: P16858 MouseSwissProt: P16858 Mouse
SwissProt: P04797 RatSwissProt: P04797 Rat

其它名称

• 38 kDa BFA-dependent ADP-ribosylation substrate antibody
• aging associated gene 9 protein antibody
• Aging-associated gene 9 protein antibody

more

其它名称其它名称

• 38 kDa BFA-dependent ADP-ribosylation substrate antibody
• aging associated gene 9 protein antibody
• Aging-associated gene 9 protein antibody

38 kDa BFA-dependent ADP-ribosylation substrate antibody

38 kDa BFA-dependent ADP-ribosylation substrate antibody

aging associated gene 9 protein antibody

aging associated gene 9 protein antibody

Aging-associated gene 9 protein antibody

Aging-associated gene 9 protein antibody moremore

应用

WB: 1:5,000-1:640,000
ICC/IF: 1:50
IHC-P: 1:50
FC: 1:50
IP: Use at an assay dependent concentration.

应用

应用

WB: 1:5,000-1:640,000
ICC/IF: 1:50
IHC-P: 1:50
FC: 1:50
IP: Use at an assay dependent concentration.

WB: 1:5,000-1:640,000
ICC/IF: 1:50
IHC-P: 1:50
FC: 1:50
IP: Use at an assay dependent concentration.

• 

  Fig1: Western blot analysis of GAPDH on Hela cell lysates with Rabbit anti-GAPDH antibody (ET1601-4).
  
  Hela cell lysates at 10 µg/Lane.
  
  Predicted band size: 36 kDa
  Observed band size: 36 kDa
  
  12% SDS-PAGE gel.
  
  Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  

  Fig2: Western blot analysis of GAPDH on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4, 1/5,000) was used in 5% NFDM/TBST at room temperature for 1 hour. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 45 mins at room temperature.
  
  Positive control:
  Lane 1: PC-3 cell lysate, 10 µg/Lane
  Lane 2: Mouse colon tissue lysate, 20 µg/Lane
  Lane 3: SH-SY5Y cell lysate, 10 µg/Lane
  Lane 4: PC-3 cell lysate, 10 µg/Lane
  Lane 5: NIH/3T3 cell lysate, 10 µg/Lane
  Lane 6: SK-Br-3 cell lysate, 10 µg/Lane
  Lane 7: Rat brain tissue lysate, 20 µg/Lane

  

  Fig3: Western blot analysis of GAPDH on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4, 1/10,000) was used in 5% NFDM/TBST at room temperature for 1 hour. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 45 mins at room temperature.
  
  Positive control:
  Lane 1: Rat liver tissue lysate, 20 µg/Lane
  Lane 2: Rat lung tissue lysate, 20 µg/Lane
  Lane 3: Rat lung tissue lysate, 20 µg/Lane
  Lane 4: Rat heart tissue lysate, 20 µg/Lane
  Lane 5: Rat cerebellum tissue lysate, 20 µg/Lane
  Lane 6: Rat skeletal muscle tissue lysate, 20 µg/Lane
  Lane 7: Rat spleen tissue lysate, 20 µg/Lane
  Lane 8: Rat small intestine tissue lysate, 20 µg/Lane

  

  Fig4: Western blot analysis of GAPDH on different lysates with Rabbit anti-GAPDH antibody (ET1601-4) at 1/80,000 dilution.
  
  Cell lysates at 10 µg/Lane, tissue lysates at 20 µg/Lane.
  
  Predicted band size: 36 kDa
  Observed band size: 36 kDa
  
  Exposure time: 1 minute;
  
  12% SDS-PAGE gel.
  
  Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4) at 1/80,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  

  Fig5: ICC staining of GAPDH in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  

  Fig6: ICC staining of GAPDH in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  

  Fig7: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1601-4, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  

  Fig8: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1601-4, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  

  Fig9: Flow cytometric analysis of GAPDH was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1601-4, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).

Fig1: Western blot analysis of GAPDH on Hela cell lysates with Rabbit anti-GAPDH antibody (ET1601-4).

Hela cell lysates at 10 µg/Lane.

Predicted band size: 36 kDa
Observed band size: 36 kDa

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of GAPDH on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4, 1/5,000) was used in 5% NFDM/TBST at room temperature for 1 hour. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 45 mins at room temperature.

Positive control:
Lane 1: PC-3 cell lysate, 10 µg/Lane
Lane 2: Mouse colon tissue lysate, 20 µg/Lane
Lane 3: SH-SY5Y cell lysate, 10 µg/Lane
Lane 4: PC-3 cell lysate, 10 µg/Lane
Lane 5: NIH/3T3 cell lysate, 10 µg/Lane
Lane 6: SK-Br-3 cell lysate, 10 µg/Lane
Lane 7: Rat brain tissue lysate, 20 µg/Lane

Fig3: Western blot analysis of GAPDH on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4, 1/10,000) was used in 5% NFDM/TBST at room temperature for 1 hour. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 45 mins at room temperature.

Positive control:
Lane 1: Rat liver tissue lysate, 20 µg/Lane
Lane 2: Rat lung tissue lysate, 20 µg/Lane
Lane 3: Rat lung tissue lysate, 20 µg/Lane
Lane 4: Rat heart tissue lysate, 20 µg/Lane
Lane 5: Rat cerebellum tissue lysate, 20 µg/Lane
Lane 6: Rat skeletal muscle tissue lysate, 20 µg/Lane
Lane 7: Rat spleen tissue lysate, 20 µg/Lane
Lane 8: Rat small intestine tissue lysate, 20 µg/Lane

Fig4: Western blot analysis of GAPDH on different lysates with Rabbit anti-GAPDH antibody (ET1601-4) at 1/80,000 dilution.

Cell lysates at 10 µg/Lane, tissue lysates at 20 µg/Lane.

Predicted band size: 36 kDa
Observed band size: 36 kDa

Exposure time: 1 minute;

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4) at 1/80,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Fig5: ICC staining of GAPDH in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Fig6: ICC staining of GAPDH in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Fig7: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1601-4, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig8: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1601-4, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig9: Flow cytometric analysis of GAPDH was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1601-4, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).

Fig1: Western blot analysis of GAPDH on Hela cell lysates with Rabbit anti-GAPDH antibody (ET1601-4).

Hela cell lysates at 10 µg/Lane.

Predicted band size: 36 kDa
Observed band size: 36 kDa

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Fig1:Fig1: Western blot analysis of GAPDH on Hela cell lysates with Rabbit anti-GAPDH antibody (ET1601-4).

Hela cell lysates at 10 µg/Lane.

Predicted band size: 36 kDa
Observed band size: 36 kDa

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of GAPDH on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4, 1/5,000) was used in 5% NFDM/TBST at room temperature for 1 hour. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 45 mins at room temperature.

Positive control:
Lane 1: PC-3 cell lysate, 10 µg/Lane
Lane 2: Mouse colon tissue lysate, 20 µg/Lane
Lane 3: SH-SY5Y cell lysate, 10 µg/Lane
Lane 4: PC-3 cell lysate, 10 µg/Lane
Lane 5: NIH/3T3 cell lysate, 10 µg/Lane
Lane 6: SK-Br-3 cell lysate, 10 µg/Lane
Lane 7: Rat brain tissue lysate, 20 µg/Lane

Fig2:Fig2: Western blot analysis of GAPDH on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4, 1/5,000) was used in 5% NFDM/TBST at room temperature for 1 hour. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 45 mins at room temperature.

Positive control:
Lane 1: PC-3 cell lysate, 10 µg/Lane
Lane 2: Mouse colon tissue lysate, 20 µg/Lane
Lane 3: SH-SY5Y cell lysate, 10 µg/Lane
Lane 4: PC-3 cell lysate, 10 µg/Lane
Lane 5: NIH/3T3 cell lysate, 10 µg/Lane
Lane 6: SK-Br-3 cell lysate, 10 µg/Lane
Lane 7: Rat brain tissue lysate, 20 µg/Lane

Fig3: Western blot analysis of GAPDH on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4, 1/10,000) was used in 5% NFDM/TBST at room temperature for 1 hour. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 45 mins at room temperature.

Positive control:
Lane 1: Rat liver tissue lysate, 20 µg/Lane
Lane 2: Rat lung tissue lysate, 20 µg/Lane
Lane 3: Rat lung tissue lysate, 20 µg/Lane
Lane 4: Rat heart tissue lysate, 20 µg/Lane
Lane 5: Rat cerebellum tissue lysate, 20 µg/Lane
Lane 6: Rat skeletal muscle tissue lysate, 20 µg/Lane
Lane 7: Rat spleen tissue lysate, 20 µg/Lane
Lane 8: Rat small intestine tissue lysate, 20 µg/Lane

Fig3:Fig3: Western blot analysis of GAPDH on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4, 1/10,000) was used in 5% NFDM/TBST at room temperature for 1 hour. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 45 mins at room temperature.

Positive control:
Lane 1: Rat liver tissue lysate, 20 µg/Lane
Lane 2: Rat lung tissue lysate, 20 µg/Lane
Lane 3: Rat lung tissue lysate, 20 µg/Lane
Lane 4: Rat heart tissue lysate, 20 µg/Lane
Lane 5: Rat cerebellum tissue lysate, 20 µg/Lane
Lane 6: Rat skeletal muscle tissue lysate, 20 µg/Lane
Lane 7: Rat spleen tissue lysate, 20 µg/Lane
Lane 8: Rat small intestine tissue lysate, 20 µg/Lane

Fig4: Western blot analysis of GAPDH on different lysates with Rabbit anti-GAPDH antibody (ET1601-4) at 1/80,000 dilution.

Cell lysates at 10 µg/Lane, tissue lysates at 20 µg/Lane.

Predicted band size: 36 kDa
Observed band size: 36 kDa

Exposure time: 1 minute;

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4) at 1/80,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Fig4:Fig4: Western blot analysis of GAPDH on different lysates with Rabbit anti-GAPDH antibody (ET1601-4) at 1/80,000 dilution.

Cell lysates at 10 µg/Lane, tissue lysates at 20 µg/Lane.

Predicted band size: 36 kDa
Observed band size: 36 kDa

Exposure time: 1 minute;

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4) at 1/80,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Fig5: ICC staining of GAPDH in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Fig5:Fig5: ICC staining of GAPDH in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Fig6: ICC staining of GAPDH in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Fig6:Fig6: ICC staining of GAPDH in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Fig7: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1601-4, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig7:Fig7: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂2O and PBS, and then probed with the primary antibody (ET1601-4, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig8: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1601-4, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig8:Fig8: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂2O and PBS, and then probed with the primary antibody (ET1601-4, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig9: Flow cytometric analysis of GAPDH was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1601-4, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).

Fig9:Fig9: Flow cytometric analysis of GAPDH was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1601-4, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).

• 背景文献

• 1. "High-resolution structure of human D-glyceraldehyde-3-phosphate dehydrogenase."Jenkins J.L., Tanner J.J.Acta Crystallogr. D 62:290-301(2006)
  
  2. "Structural analysis of human liver glyceraldehyde-3-phosphate dehydrogenase."Ismail S.A., Park H.W.Acta Crystallogr. D 61:1508-1513(2005)

背景文献

背景文献

背景文献

1. "High-resolution structure of human D-glyceraldehyde-3-phosphate dehydrogenase."Jenkins J.L., Tanner J.J.Acta Crystallogr. D 62:290-301(2006)

2. "Structural analysis of human liver glyceraldehyde-3-phosphate dehydrogenase."Ismail S.A., Park H.W.Acta Crystallogr. D 61:1508-1513(2005)

1. "High-resolution structure of human D-glyceraldehyde-3-phosphate dehydrogenase."Jenkins J.L., Tanner J.J.Acta Crystallogr. D 62:290-301(2006)

2. "Structural analysis of human liver glyceraldehyde-3-phosphate dehydrogenase."Ismail S.A., Park H.W.Acta Crystallogr. D 61:1508-1513(2005)