Overview

  • Product name
    Anti-GAPDH antibody - Loading Control
    See all GAPDH primary antibodies
  • Description
    Goat polyclonal to GAPDH - Loading Control
  • Host species
    Goat
  • Tested applications
    Suitable for: WB, ICC/IFmore 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.

Properties

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 Abreviews 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

  • Function
    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.
  • Pathway
    Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
  • Sequence similarities
    Belongs 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 localization
    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.
  • 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
    • 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

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 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : 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.
  • ab9483 staining GAPDH in HeLa by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehydeand. Samples were incubated with primary antibody (1/200) for 30 minutes. An undiluted Alexa Fluor® 568-conjugated Donkey anti-goat polyclonal was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Wallace LM  et al. Pre-clinical Safety and Off-Target Studies to Support Translation of AAV-Mediated RNAi Therapy for FSHD. Mol Ther Methods Clin Dev 8:121-130 (2018). Read more (PubMed: 29387734) »
  • Chen W  et al. HSPA12B overexpression induces cisplatin resistance in non-small-cell lung cancer by regulating the PI3K/Akt/NF-?B signaling pathway. Oncol Lett 15:3883-3889 (2018). Read more (PubMed: 29556279) »
See all 85 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Feb 21 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Sep 25 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted May 20 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
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

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

Dr. Sophie Pezet

Verified customer

Submitted Jul 31 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde

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"

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