Anti-GAPDH antibody - Loading Control (ab9483)

Overview

  • Product nameAnti-GAPDH antibody - Loading ControlSee all GAPDH primary antibodies ...
  • Description
    Goat polyclonal to GAPDH - Loading Control
  • Tested applicationsWB, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Full length native protein from human erythrocytes.

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; NIH3T3. This antibody also gave a positive signal in Human brain tissue lysate.

Applications

Our Abpromise guarantee covers the use of ab9483 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
WB 1/1000. Detects a band of approximately 37 kDa (predicted molecular weight: 35.8 kDa).
ICC/IF Use a concentration of 1 µg/ml.

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
    modifications
    S-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
  • Alternative names
    • Aging associated gene 9 protein antibody
    • G3P_HUMAN antibody
    • G3PD antibody
    • G3PD antibody
    • G3PDH antibody
    • GAPD antibody
    • GAPDH antibody
    • GAPDH antibody
    • Glyceraldehyde 3 phosphate dehydrogenase antibody
    • Glyceraldehyde 3 phosphate dehydrogenase liver antibody
    • Glyceraldehyde 3 phosphate dehydrogenase muscle antibody
    • Glyceraldehyde-3-phosphate dehydrogenase antibody
    • MGC88685 antibody
    • OCAS p38 component antibody
    • OCT1 coactivator in S phase 38 KD Component antibody
    • Peptidyl-cysteine S-nitrosylase GAPDH antibody
    see all

Anti-GAPDH antibody - Loading Control images

  • All lanes : Anti-GAPDH antibody - Loading Control (ab9483) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Rabbit polyclonal to Goat IgG (Alexa Fluor® 680) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 35.8 kDa
    Observed band size : 37 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab9483 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal donkey 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 (ab9483, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 donkey anti-goat 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa cells at 1µg/ml.

References for Anti-GAPDH antibody - Loading Control (ab9483)

This product has been referenced in:
  • Longati P  et al. 3D pancreatic carcinoma spheroids induce a matrix-rich, chemoresistant phenotype offering a better model for drug testing. BMC Cancer 13:95 (2013). WB ; Human . Read more (PubMed: 23446043) »
  • Berisha SZ  et al. Transcriptome analysis of genes regulated by cholesterol loading in two strains of mouse macrophages associates lysosome pathway and ER stress response with atherosclerosis susceptibility. PLoS One 8:e65003 (2013). WB ; Mouse . Read more (PubMed: 23705026) »

See all 21 Publications for this product

Product Wall

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (U2OS osteosarcoma)
Specification U2OS osteosarcoma
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Feb 21 2014

Application Western blot
Sample Mouse Cell lysate - whole cell (skeletal muscle)
Loading amount 50 µg
Specification skeletal muscle
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Sep 25 2012

Thank you very much for your call today and for letting us know about the trouble with this antibody.

As we discussed, I'm sending a free of charge vial from a new lot on the order *** which should arrive tomorrow.

Please keep me ud...

Read More
Application Western blot
Sample Mouse Tissue lysate - whole (aorta)
Loading amount 20 µg
Specification aorta
Gel Running Conditions Reduced Denaturing (Bis-tris 4-12%)
Blocking step Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted May 20 2010

Application Western blot
Sample Human Cell lysate - whole cell (U2OS cells)
Loading amount 50 µg
Specification U2OS cells
Gel Running Conditions Reduced Denaturing (4-20% tris-glycine)
Blocking step normal horse serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. Natalie Farny

Verified customer

Submitted Oct 23 2009

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (Brain sections)
Specification Brain sections
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

Submitted Jul 31 2009

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Username

Dr. Kirk McManus

Verified customer

Submitted Oct 03 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"