Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)

Overview

• Product nameAnti-GAPDH antibody [mAbcam 9484] - Loading ControlSee all GAPDH primary antibodies ...
• Description
  Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control
• SpecificityThis antibody shows a single clean band in Western blotting on cell lysates (rat and chinese hamster lysates also show another slightly lower band).
• Tested applicationsICC/IF, WB, IHC-P, Flow Cyt more details
• Species reactivity
  Reacts with: Mouse, Rat, Rabbit, Chicken, Cow, Dog, Human, Pig, Xenopus laevis, Chinese Hamster
  
• Immunogen

  Full length native protein from human erythrocytes.

• General notes

  We can conjugate this antibody to FITC for you (please see ab150196 for details).

Properties

• FormLiquid
• Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: 0.01% Sodium Azide
  Constituents: PBS, pH 7.4
• Concentration information loading...
• PurityIgG fraction
• Clonality Monoclonal
• Clone numbermAbcam 9484
• MyelomaSp2/0-Ag14
• IsotypeIgG2b
• Light chain typekappa
• Research Areas
  • Metabolism
  • Types of disease
  • Neurodegenerative disease

  • Metabolism
  • Types of disease
  • Cancer

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

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

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

  • Signal Transduction
  • Metabolism
  • Energy Metabolism

  • Isotype/Loading Controls
  • Loading Controls
  • GAPDH

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

  • Neuroscience
  • Neurology process
  • Neurodegenerative disease
  • Huntington's disease

Applications

Target

• FunctionHas 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.
• PathwayCarbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
• Sequence similaritiesBelongs to the glyceraldehyde-3-phosphate dehydrogenase family.
• Post-translational
  modificationsS-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
  ISGylated.
• Cellular localizationCytoplasm > 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.

Target information above from: UniProt accession P04406 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 374193 Chicken
  • Entrez Gene: 2597 Human
  • Entrez Gene: 100042025 Mouse
  • Entrez Gene: 14433 Mouse
  • Entrez Gene: 100009074 Rabbit
  • Entrez Gene: 24383 Rat
  • Entrez Gene: 685186 Rat
  • Entrez Gene: 380259 Xenopus laevis

  • Omim: 138400 Human
  • SwissProt: P00356 Chicken
  • SwissProt: P04406 Human
  • SwissProt: P16858 Mouse
  • SwissProt: P00355 Pig
  • SwissProt: P46406 Rabbit
  • SwissProt: P04797 Rat
  • SwissProt: P51469 Xenopus laevis
  • Unigene: 544577 Human
  • Unigene: 592355 Human
  • Unigene: 598320 Human
  • Unigene: 304088 Mouse
  • Unigene: 309092 Mouse
  • Unigene: 317779 Mouse
  • Unigene: 343110 Mouse
  • Unigene: 392463 Mouse
  • Unigene: 392480 Mouse
  • Unigene: 414470 Mouse
  • Unigene: 458138 Mouse
  • Unigene: 458416 Mouse
  • Unigene: 475698 Mouse
  • Unigene: 129558 Rat
  • Unigene: 91450 Rat
  • Unigene: 995 Xenopus laevis

  see all

• Alternative names
  Aging associated gene 9 protein antibodyG3P_HUMAN antibodyG3PD antibody

  G3PD antibodyG3PDH antibodyGAPD antibodyGAPDH antibodyGAPDH antibodyGlyceraldehyde 3 phosphate dehydrogenase antibodyGlyceraldehyde 3 phosphate dehydrogenase liver antibodyGlyceraldehyde 3 phosphate dehydrogenase muscle antibodyGlyceraldehyde-3-phosphate dehydrogenase antibodyMGC88685 antibodyOCAS p38 component antibodyOCT1 coactivator in S phase 38 KD Component antibodyPeptidyl-cysteine S-nitrosylase GAPDH antibody

  see all

Anti-GAPDH antibody [mAbcam 9484] - Loading Control images

• 

  Western blot - GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
  All lanes : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/5000 dilution
  
  Lane 1 : Hela whole cell (Human)
  Lane 2 : 3T3 cell (Mouse)
  Lane 3 : Rat brain
  Lane 4 : Xenopus embryo
  Lane 5 : Chicken Liver
  Lane 6 : EBTr cell (Cow)
  Lane 7 : CHO cell (Chinese hamster)
  Lane 8 : Pig liver
  
  Secondary
  Rabbit polyclonal Secondary Antibody to Mouse IgG - H&L (HRP) (ab6728) at 1/5000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size : 40.2 kDa
  
  
  Exposure time : 10 seconds
  
  The membrane was blocked in 5%BSA in TBST for one hour, then incubated for one hour in primary antibody diluted in TBST.
• 

  Western blot - GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
  Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 0.5 µg/ml + HeLa cell lysate
  
  Secondary
  Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP) (ab6789) at 1/5000 dilution
  developed using the ECL technique
  
  Performed under non-reducing conditions.
  
  Predicted band size : 40.2 kDa
  
  
  Exposure time : 30 seconds
• 

  Western blot - GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
  Lanes 1 - 5 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/1000 dilution (Blocked in 5% MILK)
  Lanes 6 - 10 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/1000 dilution (Blocked in 5% BSA)
  
  Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
  Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
  Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)
  Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)
  Lane 5 : HEK293 Human embryonic kidney cell line Whole Cell Lysate (ab7902)
  Lane 6 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
  Lane 7 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
  Lane 8 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)
  Lane 9 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)
  Lane 10 : HEK293 Human embryonic kidney cell line Whole Cell Lysate (ab7902)
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  Goat anti-Mouse (HRP conjugated) at 1/5000 dilution
  
  Predicted band size : 40.2 kDa
  Observed band size : 40.2 kDa
  
• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
  IHC image of GAPDH staining in human liver FFPE section, performed on a Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9484, 5µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
• 

  Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
  ICC/IF image of ab9484 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9484, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  
  
• 

  Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
  ICC/IF image of ab9484 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9484, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  
  
• 

  Flow Cytometry - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)Image courtesy of Dr Mahesh Shivananjappa by Abreview.
  Flow Cytometry analysis of GAPDH using ab9484.
  
  Human monocytes were fixed in paraformaldehyde and permeabilized. ab9484 was used at a 1/200 dilution. The secondary used was an Alexa-Fluor 488 conjugated chicken anti-rabbit IgG (H+L) polyclonal, used at a 1/500 dilution.

  See Abreview

• 

  Flow Cytometry - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
  Overlay histogram showing HeLa cells stained with ab9484 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab9484, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.