Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
![Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)](https://www.abcam.com/ps/products/9/ab9484/Images/ab9484-344444-DS4ab9484AP41529711m.jpg "Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)")
Key features and details
- Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control
- Suitable for: WB, IHC-P, Flow Cyt
- Reacts with: Mouse, Rat, Chicken, Cow, Human, Pig, Xenopus laevis, Chinese hamster
- Isotype: IgG2b
Overview
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Product name
Anti-GAPDH antibody [mAbcam 9484] - Loading Control
See all GAPDH primary antibodies -
Description
Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control -
Host species
Mouse -
Specificity
In western blot, this product typically gives a lower signal in rat lysates compared to human and mouse lysates.
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Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanIHC-P HumanWB MouseRatChickenCowHumanPigXenopus laevisChinese hamster
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Immunogen
Full length native protein (purified) corresponding to Human GAPDH.
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Positive control
- WB: A431, Jurkat, HEK293, HeLa, NIH 3T3, PC-12, EBTr and CHO cell lysates; Rat brain, chicken and pig liver and Xenopus laevis embryo tissue lysates. IHC-P: Human liver tissue. Flow Cyt: HeLa cells
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General notes
For Western blotting, do not use milk for blocking. Our labs have extensively tested the blocking conditions for this antibody and recommend using 5% BSA for 1 hour. The comparison data is shown in the images section.
This antibody clone [mAbcam 9484] is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading... -
Purity
IgG fraction -
Clonality
Monoclonal -
Clone number
mAbcam 9484 -
Myeloma
Sp2/0-Ag14 -
Isotype
IgG2b -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
- Anti-GAPDH antibody [mAbcam 9484] - BSA and Azide free (ab174646)
- Alexa Fluor® 680 Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab184095)
- Alexa Fluor® 790 Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab184578)
- Alexa Fluor® 647 Anti-GAPDH antibody [mAbcam 9484] (ab196697)
- HRP Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9482)
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab9484 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| Flow Cyt |
Human
|
| IHC-P |
Human
|
| WB |
Mouse
Rat
Chicken
Cow
Human
Pig
Xenopus laevis
Chinese hamster
|
| All applications |
Rabbit
Dog
Cynomolgus monkey
|
| Application | Abreviews | Notes |
|---|---|---|
| WB | (52) |
Use a concentration of 0.1 - 1 µg/ml. Predicted molecular weight: 36 kDa.
Do not block with milk. Block with 5% BSA for 1 hour. Our labs have thoroughly investigated the blocking conditions for this antibody. We found that milk significantly decreases the signal and is therefore not a suitable blocking agent for this antibody (see images). |
| IHC-P |
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
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| Flow Cyt | (1) |
Use 1µg for 106 cells.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. We recommend Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879) secondary antibody |
| Notes |
|---|
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WB
Use a concentration of 0.1 - 1 µg/ml. Predicted molecular weight: 36 kDa. Do not block with milk. Block with 5% BSA for 1 hour. Our labs have thoroughly investigated the blocking conditions for this antibody. We found that milk significantly decreases the signal and is therefore not a suitable blocking agent for this antibody (see images). |
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IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
|
Flow Cyt
Use 1µg for 106 cells. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. We recommend Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879) secondary antibody |
Target
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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
modificationsS-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
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Database links
- Entrez Gene: 374193 Chicken
- Entrez Gene: 281181 Cow
- Entrez Gene: 403755 Dog
- Entrez Gene: 2597 Human
- Entrez Gene: 100042025 Mouse
- Entrez Gene: 14433 Mouse
- Entrez Gene: 396823 Pig
- Entrez Gene: 100009074 Rabbit
see all -
Alternative names
- 38 kDa BFA-dependent ADP-ribosylation substrate antibody
- aging associated gene 9 protein antibody
- Aging-associated gene 9 protein antibody
see all
Images
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Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)All lanes : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1 µg/ml
Lane 1 : HeLa Whole Cell Lysate
Lane 2 : NIH 3T3 Whole Cell Lysate
Lane 3 : PC12 Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 36 kDa
Exposure time: 1 minuteThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab9484 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)](https://www.abcam.com/ps/products/9/ab9484/Images/ab9484-17137-anti-gapdh-antibody-mabcam-9484-loading-control-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)IHC image of GAPDH staining in human liver FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9484, 5µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. -
![Flow Cytometry - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)](https://www.abcam.com/ps/products/9/ab9484/Images/ab9484-17133-anti-gapdh-antibody-mabcam-9484-loading-control-flow-cytometry.jpg)
Flow Cytometry - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)Overlay histogram showing HeLa cells stained with ab9484 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab9484, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was goat anti-mouse DyLight® 488 (IgG H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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![Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)](https://www.abcam.com/ps/products/9/ab9484/Images/ab9484-17140-anti-gapdh-antibody-mabcam-9484-loading-control-western-blot.jpg)
Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)All lanes : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/5000 dilution
Lane 1 : Hela whole cell (Human)
Lane 2 : 3T3 cell (Mouse)
Lane 3 : Rat brain
Lane 4 : Xenopus laevis embryo
Lane 5 : Chicken Liver
Lane 6 : EBTr cell (Cow)
Lane 7 : CHO cell (Chinese hamster)
Lane 8 : Pig liver
Secondary
All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsThe membrane was blocked in 5% BSA in TBST for 1 hour, then incubated for 1 hour in primary antibody diluted in TBST.
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![Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)](https://www.abcam.com/ps/products/9/ab9484/Images/ab9484-17138-anti-gapdh-antibody-mabcam-9484-loading-control-western-blot.jpg)
Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)Lanes 1-5 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/1000 dilution (Blocked in 5% milk)
Lanes 6-10 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/1000 dilution (Blocked in 5% BSA)
Lanes 1 & 6 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lanes 2 & 7 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lanes 3 & 8 :A-431 whole cell lysate (ab7909)
Lanes 4 & 9 :Jurkat whole cell lysate (ab7899)
Lanes 5 & 10 :HEK-293 whole cell lysate (ab7902)
Lysates/proteins at 20 µg per lane.
Secondary
Lanes 1-5 : Goat anti-Mouse (HRP conjugated) at 1/5000 dilution
Lanes 6-10 : Goat anti-Mouse (HRP conjugated) at 1/5000 dilution
Predicted band size: 36 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?The membrane 1-5 was blocked in 5% milk (1 hour). The membrane 6-10 was blocked in 5% BSA (1 hour). Milk is not a suitable blocking agent and significantly decreases the signal on the membrane.
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![Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)](https://www.abcam.com/ps/products/9/ab9484/Images/ab9484-17139-anti-gapdh-antibody-mabcam-9484-loading-control-western-blot.jpg)
Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 0.5 µg/ml + HeLa cell lysate
Secondary
Goat Anti-Mouse IgG H&L (HRP) (ab6789) at 1/5000 dilution
Developed using the ECL technique.
Performed under non-reducing conditions.
Predicted band size: 36 kDa
Exposure time: 30 seconds
Datasheets and documents
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SDS download
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Datasheet download
References (614)
ab9484 has been referenced in 614 publications.
- Spadotto V et al. PRMT1-mediated methylation of the microprocessor-associated proteins regulates microRNA biogenesis. Nucleic Acids Res 48:96-115 (2020). PubMed: 31777917
- Venugopal N et al. The primary cilium dampens proliferative signaling and represses a G2/M transcriptional network in quiescent myoblasts. BMC Mol Cell Biol 21:25 (2020). PubMed: 32293249
- Chu Y et al. Argonaute binding within 3'-untranslated regions poorly predicts gene repression. Nucleic Acids Res N/A:N/A (2020). PubMed: 32501500
- Murphy-Royal C et al. Stress gates an astrocytic energy reservoir to impair synaptic plasticity. Nat Commun 11:2014 (2020). PubMed: 32332733
- Schumann U et al. Multiple links between 5-methylcytosine content of mRNA and translation. BMC Biol 18:40 (2020). PubMed: 32293435