Anti-GAPDH antibody - Loading Control (HRP) (ab9385)

Overview

  • Product nameAnti-GAPDH antibody - Loading Control (HRP)
    See all GAPDH primary antibodies
  • Description
    Rabbit polyclonal to GAPDH - Loading Control (HRP)
  • ConjugationHRP
  • Conjugation notesThis antibody has been conjugated with HRP. The concentration of the original antibody was 1 mg/ml, following conjugation the concentration observed will be higher.
  • Tested applicationsWBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Saccharomyces cerevisiae, Xenopus laevis
  • Immunogen

    Full length native protein from human erythrocytes.

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; Jurkat; A431; Hek293.
  • General notes

     

    This antibody is the HRP conjugated version of ab9485, for more convenient use as a loading control antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab9385 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 1/5000. Detects a band of approximately 38 kDa (predicted molecular weight: 36 kDa). Milk blocking may cause 'no bands' problem. Please try BSA as well.

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
    • 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 (HRP) images

  • Lanes 1 - 4 : Anti-GAPDH antibody - Loading Control (HRP) (ab9385) at 1/5000 dilution ((Blocked in 5% BSA))
    Lanes 5 - 8 : Anti-GAPDH antibody - Loading Control (HRP) (ab9385) at 1/5000 dilution ((Blocked in 5% MILK))

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab52254)
    Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)
    Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate (ab7902)
    Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)
    Lane 6 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab52254)
    Lane 7 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)
    Lane 8 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate (ab7902)

    Lysates/proteins at 10 µg per lane.

    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 36 kDa
  • All lanes : Anti-GAPDH antibody - Loading Control (HRP) (ab9385) at 1 µg/ml

    Lane 1 : HeLa cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : 293 cell lysate

    Lysates/proteins at 20 µg per lane.


    Performed under reducing conditions.

    Predicted band size : 36 kDa
    Observed band size : 38 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

References for Anti-GAPDH antibody - Loading Control (HRP) (ab9385)

This product has been referenced in:
  • Rottmar M  et al. Interference with the contractile machinery of the fibroblastic chondrocyte cytoskeleton induces re-expression of the cartilage phenotype through involvement of PI3K, PKC and MAPKs. Exp Cell Res 320:175-87 (2014). WB ; Cow . Read more (PubMed: 24246223) »
  • Schroeder EA & Shadel GS Crosstalk between mitochondrial stress signals regulates yeast chronological lifespan. Mech Ageing Dev 135:41-9 (2014). Saccharomyces cerevisiae . Read more (PubMed: 24373996) »

See all 20 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing (Acrylamide gel, 10%)
Sample Human Tissue lysate - whole (brain (frontal cortex))
Specification brain (frontal cortex)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 31 2013

Thank you for contacting us.
I'm sorry to hear that the antibody is not working as expected.

As we discussed over the phone, the following suggestions might help with improving your results:

1) increase primary antibody concen...

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Thank you for confirming these details and for your cooperation.
As requested, I have issued a free of charge replacement with ab9482. The order number is 1130988.
To check the status of the order please contact our Customer Service team and ...

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Unfortunately we currently do not have an alternative lot for ab9385, however we have an alternative antibody, reference ab9482, www.abcam.com/ab9482.
Would you like to try the other reference catalog we have (ab9482)?
I can still send you ano...

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Thank you for contacting us. I am sorry to hear this antibody is not providing satisfactory results.
The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I apprecia...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

In or...

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Thank you for your enquiry and your interest in our product.

We do not test our antibodies in Candida albicans on a regular basis, so it is not known if any of them would work in this species. However, the most popular loading controls in mam...

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Thank you for your inquiry. I heard back from the lab that the immunogen for ab9385 is a recombinant protein and rabbits were immunized with this recombinant sourced from Sigma (G6019). As this was not performed in-house we have no details about how...

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Application Western blot
Sample African Green Monkey Cell lysate - whole cell (CV1 cell lysate)
Loading amount 10 µg
Specification CV1 cell lysate
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Mr. Nir Drayman

Verified customer

Submitted Jan 12 2011

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