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

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Overview

Product name

Anti-GAPDH antibody (HRP) - Loading Control
See all GAPDH products (9) ...

Description

Rabbit polyclonal to GAPDH - Loading Control (HRP)

Conjugation

HRP

Conjugation notes

This 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 applications

WBmore details

Cross reactivity

Reacts with

Mouse, Rat, 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.

Properties

Form

Liquid

Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.

Storage buffer

Preservative: None
Constituents: 50% Glycerol, PBS, pH 7.4

Concentration

Concentration information loading...

Purity

Immunogen affinity purified

Primary antibody notes

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

Clonality

Polyclonal

Isotype

IgG

  • Western blot - GAPDH antibody (HRP) - Loading Control (ab9385)Western blot - GAPDH antibody (HRP) - Loading Control (ab9385) image (enlarge)

  • Western blot - GAPDH antibody (ab9385)Western blot - GAPDH antibody (ab9385) image (enlarge)

Applications

Show applications key

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.

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.

Target information above from: UniProt accessionP04406 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).

Information by UniProt

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 (HRP) - Loading Control images:

  Western blot - GAPDH antibody (HRP) - Loading Control (ab9385)

Western blot - GAPDH antibody (HRP) - Loading Control (ab9385)

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


The membrane 1-4 was blocked in 5% BSA (1 hour). The membrane 5-8 was blocked in 5% MILK (1 hour). Abcam routinely uses 5% BSA to block, however following recent customer feedback our labs investigated the effect of 5% milk blocking. We can now confirm that milk is not a suitable blocking agent for this antibody and significantly decreases the signal on the membrane.

  Western blot - GAPDH antibody (ab9385)

Western blot - GAPDH antibody (ab9385)

All lanes : Anti-GAPDH antibody (HRP) - Loading Control (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 (HRP) - Loading Control (ab9385)

This product has been referenced in:

  • Kim JYet al. Low molecular weight silk fibroin increases alkaline phosphatase and type I collagen expression in MG63 cells. BMB Rep 43:52-6 (2010). WB.Read more (PubMed: 20132736) »
  • Heidrich FMet al. Chromogranin B regulates calcium signaling, nuclear factor kappaB activity, and brain natriuretic peptide production in cardiomyocytes. Circ Res 102:1230-8 (2008). WB; Mouse.Read more (PubMed: 18420944) »

See all 9 publications for this product

Publishing research using ab9385? Please let us know so that we can cite the reference in this datasheet

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