Anti-GAPDH antibody - Loading Control (ab9483)
- Datasheet
- References (85)
- Protocols
Overview
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Product nameAnti-GAPDH antibody - Loading Control
See all GAPDH primary antibodies -
DescriptionGoat polyclonal to GAPDH - Loading Control
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Host speciesGoat
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Tested applicationsSuitable for: WB, ICC/IFmore details
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Species reactivityReacts with: Mouse, Human
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Immunogen
Full length native protein from human erythrocytes.
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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
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FormLiquid
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Storage instructionsShipped 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.
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Storage bufferPreservative: 0.02% Sodium azide
This product may contain up to 3% BSA depending on the batch. For specific batch formulations please contact us. -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
- Anti-SDHB antibody [21A11AE7] (ab14714)
- Anti-SDHA antibody [2E3GC12FB2AE2] (ab14715)
- GAPDH ELISA Kit (ab176642)
- Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)
- Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)
- Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
- Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
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
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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.
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PathwayCarbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
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Sequence similaritiesBelongs to the glyceraldehyde-3-phosphate dehydrogenase family.
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Post-translational
modificationsS-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
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Database links
- Entrez Gene: 2597 Human
- Entrez Gene: 100042025 Mouse
- Entrez Gene: 14433 Mouse
- Omim: 138400 Human
- SwissProt: P04406 Human
- SwissProt: P16858 Mouse
- Unigene: 544577 Human
- Unigene: 592355 Human
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 - Loading Control (ab9483)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? -
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody - Loading Control (ab9483)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. -
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody - Loading Control (ab9483)This image is courtesy of an Abreview submitted by Kirk McManusab9483 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.
Datasheets and documents
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) »
