Overview

  • Product nameAnti-GAPDH antibodySee all GAPDH primary antibodies ...
  • Description
    Rabbit polyclonal to GAPDH
  • Tested applicationsICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Baboon
    Predicted to work with: Sheep, Rabbit, Horse, Chicken, Guinea pig, Cow, Cat, Dog, Turkey, Pig, Duck
  • Immunogen

    Synthetic peptide:

    DAPMFVMGVNHFKYA

    , corresponding to amino acids 126-140 of Human GAPDH.

  • Positive control
    • Rat heart protein extract. This antibody gave a positive result in IF in the following Formaldehyde fixed cell line: MCF7

Applications

Our Abpromise guarantee covers the use of ab22555 in the following tested applications.

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

Application Abreviews Notes
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 0.1 µg/ml. Detects a band of approximately 36 kDa.

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.
  • Information by UniProt
  • Database links
  • Alternative names
    • 38 kDa BFA-dependent ADP-ribosylation substrate antibody
    • aging associated gene 9 protein antibody
    • Aging-associated gene 9 protein antibody
    • BARS-38 antibody
    • cb609 antibody
    • EC 1.2.1.12 antibody
    • Epididymis secretory sperm binding protein Li 162eP antibody
    • G3P_HUMAN antibody
    • G3PD antibody
    • G3PDH antibody
    • GAPD antibody
    • GAPDH antibody
    • GAPDH antibody
    • Glyceraldehyde 3 phosphate dehydrogenase antibody
    • Glyceraldehyde-3-phosphate dehydrogenase antibody
    • HEL-S-162eP antibody
    • KNC-NDS6 antibody
    • MGC102544 antibody
    • MGC102546 antibody
    • MGC103190 antibody
    • MGC103191 antibody
    • MGC105239 antibody
    • MGC127711 antibody
    • MGC88685 antibody
    • OCAS, p38 component antibody
    • OCT1 coactivator in S phase, 38-KD component antibody
    • peptidyl cysteine S nitrosylase GAPDH antibody
    • Peptidyl-cysteine S-nitrosylase GAPDH antibody
    • wu:fb33a10 antibody
    see all

Anti-GAPDH antibody images

  • Anti-GAPDH antibody (ab22555) at 0.1 µg/ml + Rat heart protein extract

    Performed under reducing conditions.

    Observed band size : 36 kDa (why is the actual band size different from the predicted?)
  • All lanes : Anti-GAPDH antibody (ab22555) at 1/5000 dilution

    Lane 1 : Mouse Tissue Lysate- Whole brain
    Lane 2 : Mouse Tissue Lysate- Whole brain
    Lane 3 : Mouse Tissue Lysate- Whole brain
    Lane 4 : Mouse Tissue Lysate- Whole brain
    Lane 5 : Mouse Tissue Lysate- Whole brain
    Lane 6 : Mouse Tissue Lysate- Whole brain
    Lane 7 : Mouse Tissue Lysate- Whole brain
    Lane 8 : Mouse Tissue Lysate- Whole brain

    Lysates/proteins at 50 µg per lane.

    This image is courtesy of an anonymous Abreview

    All Lanes: ab22555 (1/15000) staining GAPDH in Mouse brain whole tissue lysate (50 µg).
    Blocking Step: 5% Milk for 1 hour at 20°C.
    Secondary Antibody: HRP-conjugated Goat anti-rabbit IgG polyclonal (1/30000).

    See Abreview

  • ab22555 stained MCF-7 cells. The cells were 4% formaldehyde fixed (10 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 ab22555 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.

References for Anti-GAPDH antibody (ab22555)

This product has been referenced in:
  • van Blitterswijk M  et al. Characterization of FUS mutations in amyotrophic lateral sclerosis using RNA-Seq. PLoS One 8:e60788 (2013). WB ; Human . Read more (PubMed: 23577159) »
  • Finkensieper A  et al. The 5-lipoxygenase pathway regulates vasculogenesis in differentiating mouse embryonic stem cells. Cardiovasc Res 86:37-44 (2010). Read more (PubMed: 19965960) »

See all 3 Publications for this product

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Application Western blot
Sample Human Tissue lysate - whole (brain)
Loading amount 50 µg
Specification brain
Gel Running Conditions Reduced Denaturing (10%SDS gel)
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 Apr 30 2010

Application Western blot
Sample Mouse Tissue lysate - whole (brain)
Loading amount 50 µg
Specification brain
Gel Running Conditions Reduced Denaturing (10%)
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 Apr 15 2010

Application Western blot
Sample Baboon Tissue lysate - whole (brain)
Loading amount 50 µg
Specification brain
Gel Running Conditions Reduced Denaturing (10% gel)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Apr 08 2010

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