• 经测试应用WB, IHC-P, ICC/IF, Flow Cytmore details
• 种属反应性
  与反应: Mouse, Rat, Chicken, Cow, Dog, Human, Monkey
• 免疫原

  Recombinant fragment within Mouse GAPDH aa 1-150. The exact sequence is proprietary.
  Database link: P16858

  Run BLAST with Run BLAST with 
• 阳性对照
  • WB: HeLa, MDBK, COS-1, MDCK, UMNSAH/DF-1, Jurkat, C6 and NIH/3T3 whole cell lysates; Mouse brain and heart lysates; Rat brain, heart, kidney and spleen lysates; Human fetal brain, heart and kidney lysates. IHC-P: Human transitional cell carcinoma of bladder, Mouse spleen and Rat spleen tissues. ICC/IF: HeLa cells. Flow: Jurkat cells.
• 常规说明

  This product is a recombinant rabbit monoclonal antibody.

  Produced using Abcam’s RabMAb^® technology. RabMAb^® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

  Alternative versions available:

  Anti-GAPDH antibody (Alexa Fluor^® 488) [EPR16884] (ab204276)

  Anti-GAPDH antibody (Alexa Fluor^® 647) [EPR16884] (ab204480)

性能

• 形式Liquid
• 存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
• 存储溶液Preservative: 0.01% Sodium azide
  Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
• 浓度100 µl 浓度为 1.485 mg/ml 40 µl 浓度为 1.485 mg/ml 10 µl 浓度为 1.485 mg/ml 
  
• 纯度Protein A purified
• 克隆单克隆
• 克隆编号EPR16884
• 同种型IgG
• 研究领域
  • Neuroscience
   
  • Neurology process
   
  • Neurodegenerative disease
  • Huntington's disease

  • Neuroscience
   
  • Neurology process
   
  • Neurodegenerative disease
  • Alzheimer's disease
   
  • Other

  • Isotype/Loading Controls
   
  • Loading Controls
   
  • GAPDH

  • Signal Transduction
   
  • Metabolism
   
  • Energy Metabolism

  • Cancer
   
  • Cancer Metabolism
   
  • Metabolic signaling pathway
  • Metabolism of carbohydrates

  • Metabolism
   
  • Pathways and Processes
   
  • Metabolic signaling pathways
  • Carbohydrate metabolism

  • Metabolism
   
  • Pathways and Processes
   
  • Metabolic signaling pathways
  • Energy transfer pathways
   
  • Energy Metabolism

  • Metabolism
   
  • Types of disease
   
  • Neurodegenerative disease

应用

靶标

• 功能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 (By similarity). 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.
• 通路Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
• 序列相似性Belongs to the glyceraldehyde-3-phosphate dehydrogenase family.
• 翻译后修饰S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
  ISGylated.
• 细胞定位Cytoplasm > cytosol. Nucleus. Cytoplasm > perinuclear region. Membrane. Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions.

• Anti-GAPDH antibody [EPR16884] 图像

  • 

    Western blot - Anti-GAPDH antibody [EPR16884] (ab181603)
    All lanes : Anti-GAPDH antibody [EPR16884] (ab181603) at 1/10000 dilution
    
    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
    Lane 2 : MDBK (Bovine kidney cell line) whole cell lysates
    Lane 3 : COS-1 (African green monkey kidney fibroblast-like cell line) whole cell lysates
    Lane 4 : MDCK (Canine kidney cell line) whole cell lysates
    
    Lysates/proteins at 20 µg per lane.
    
    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
    
    Predicted band size : 36 kDa
    Observed band size : 36 kDa
    

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • 

    Western blot - Anti-GAPDH antibody [EPR16884] (ab181603)
    All lanes : Anti-GAPDH antibody [EPR16884] (ab181603) at 1/50000 dilution
    
    Lane 1 : UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates
    Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates
    
    Lysates/proteins at 20 µg per lane.
    
    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
    
    Predicted band size : 36 kDa
    Observed band size : 36 kDa
    

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • 

    Western blot - Anti-GAPDH antibody [EPR16884] (ab181603)
    All lanes : Anti-GAPDH antibody [EPR16884] (ab181603) at 1/10000 dilution
    
    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Rat brain lysate
    Lane 4 : Rat heart lysate
    Lane 5 : Rat kidney lysate
    Lane 6 : Rat spleen lysate
    Lane 7 : C6 (Rat glial tumor cells) whole cell lysates
    Lane 8 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates
    
    Lysates/proteins at 10 µg per lane.
    
    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
    
    Predicted band size : 36 kDa
    Observed band size : 36 kDa
    

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • 

    Western blot - Anti-GAPDH antibody [EPR16884] (ab181603)
    All lanes : Anti-GAPDH antibody [EPR16884] (ab181603) at 1/10000 dilution
    
    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate
    
    Lysates/proteins at 10 µg per lane.
    
    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
    
    Predicted band size : 36 kDa
    Observed band size : 36 kDa
    

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • 

    Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16884] (ab181603)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling GAPDH with ab181603 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm and nuclear staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected withab7291 (anti-Tubulin mouse mAb) at 1/500 dilution andab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows;
    1. ab181603 at 1/250 dilution followed by ab150120(AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

  • 

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] (ab181603)

    Immunohistochemical analysis of paraffin-embedded Human transitional cell carcinoma of bladder tissue labeling GAPDH with ab181603 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic and nucleus staining on the tumor cells of transitional cell carcinoma of Human bladder is observed. Counter stained with Hematoxylin.

     

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • 

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] (ab181603)

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling GAPDH with ab181603 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocytes of mouse spleen is observed. Counter stained with Hematoxylin.

     

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • 

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] (ab181603)

    Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling GAPDH with ab181603 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocyte of rat spleen is observed. Counter stained with Hematoxylin.

     

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • 

    Flow Cytometry - Anti-GAPDH antibody [EPR16884] (ab181603)

    Flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling GAPDH with ab181603 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.