Overview

  • Product name
    Anti-GAPDH antibody - Loading Control (HRP)
    See all GAPDH primary antibodies
  • Description
    Rabbit polyclonal to GAPDH - Loading Control (HRP)
  • Host species
    Rabbit
  • Conjugation
    HRP
  • Tested applications
    Suitable for: WBmore 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.

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

  • 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 (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


    Lane 1 - 4 : GAPDH antibody - Loading Control (ab9385) at 1 ug/ml

    Lane 1 : HeLa cell lysate at 20 ug
    Lane 2 : A431 cell lysate at 20 ug
    Lane 3 : Jurkat cell lysate at 20 ug
    Lane 4 : 293 cell lysate at 20 ug

    Performed under reducing conditions.

    Exposure time : 1 minute

  • 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))

    Lanes 1 & 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)
    Lanes 2 & 6 : Jurkat (Human) Whole Cell Lysate (ab52254)
    Lanes 3 & 7 : A431 whole cell lysate (ab7909)
    Lanes 4 & 8 : HEK293 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.

References

This product has been referenced in:
  • Hu J  et al. TRPS1 Suppresses Breast Cancer Epithelial-mesenchymal Transition Program as a Negative Regulator of SUZ12. Transl Oncol 11:416-425 (2018). WB ; Human . Read more (PubMed: 29471243) »
  • Ghézali G  et al. Connexin 30 controls astroglial polarization during postnatal brain development. Development 145:N/A (2018). Read more (PubMed: 29475972) »
See all 51 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Application
Western blot
Sample
Baboon Tissue lysate - whole (Fetal skeletal muscle)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
20 µg
Specification
Fetal skeletal muscle
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Soon-Ok Kim

Verified customer

Submitted Feb 09 2017

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

Abcam user community

Verified customer

Submitted Jul 31 2013

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

Mr. Nir Drayman

Verified customer

Submitted Jan 12 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (human whole cell lysate)
Loading amount
10 µg
Specification
human whole cell lysate
Gel Running Conditions
Reduced Denaturing (4-12% PA gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Mr. Nir Drayman

Verified customer

Submitted Jan 12 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (HeLa)
Loading amount
50 µg
Specification
HeLa
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Dr. Ioannis Gavvovidis

Verified customer

Submitted Dec 15 2010

Answer

Thank you for letting me know the result of the troubleshooting tip. I am glad to hear that changing to BSA blocking solution helped to overcome the initial 'no bands' with milk blocking solution.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (lymphocytes)
Loading amount
10 µg
Specification
lymphocytes
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Dec 17 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HFFs)
Loading amount
1000 cells
Specification
HFFs
Treatment
treated with SiRNAs to various cellular proteins
Gel Running Conditions
Non-reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Dec 12 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver and Testes whole and nuclear lysates)
Loading amount
50 µg
Specification
Liver and Testes whole and nuclear lysates
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Mr. Peter Thompson

Verified customer

Submitted Jan 24 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (splenocytes)
Loading amount
150000 cells
Specification
splenocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%

Abcam user community

Verified customer

Submitted Jan 11 2007

1-10 of 12 Abreviews or Q&A

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