Recombinant Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
![Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)](https://www.abcam.com/ps/products/181/ab181603/Images/ab181603-230548-anti-gapdh-antibody-epr16884-loading-control-western-blot.jpg "Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16884] to GAPDH - Loading Control
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Chicken, Cow, Dog, Human, African green monkey
Related conjugates and formulations
Overview
-
Product name
Anti-GAPDH antibody [EPR16884] - Loading Control
See all GAPDH primary antibodies -
Description
Rabbit monoclonal [EPR16884] to GAPDH - Loading Control -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Chicken, Cow, Dog, Human, African green monkey
Predicted to work with: Monkey
-
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: HeLa, MDBK, COS-1, MDCK, UMNSAH/DF-1, Jurkat, C6 and NIH/3T3 whole cell lysates; Mouse brain and heart lysates; Rat brain, heart, kidney and spleen lysates; Human fetal brain, heart and kidney lysates. IHC-P: Human transitional cell carcinoma of bladder, Mouse spleen and Rat spleen tissues. ICC/IF: HeLa cells. Flow: Jurkat cells.
-
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16884 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
- Anti-GAPDH antibody [EPR16884] - BSA and Azide free (ab199554)
- Alexa Fluor® 488 Anti-GAPDH antibody [EPR16884] (ab204276)
- Alexa Fluor® 647 Anti-GAPDH antibody [EPR16884] (ab204480)
- HRP Anti-GAPDH antibody [EPR16884] - Loading Control (ab204481)
- Alexa Fluor® 594 Anti-GAPDH antibody [EPR16884] (ab206373)
- Alexa Fluor® 555 Anti-GAPDH antibody [EPR16884] (ab206629)
- PE Anti-GAPDH antibody [EPR16884] (ab209037)
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab181603 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 |
|---|---|---|
| Flow Cyt (Intra) |
1/200.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
| WB | (3) |
1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
|
| IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
| ICC/IF | (3) |
1/250.
|
| Notes |
|---|
|
Flow Cyt (Intra)
1/200. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
WB
1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa). |
|
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
ICC/IF
1/250. |
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
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
-
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: 24383 Rat
- Entrez Gene: 685186 Rat
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
-
Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)All lanes : Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603) at 1/10000 dilution
Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
Lane 2 : MDBK (Bovine kidney cell line) whole cell lysates
Lane 3 : COS-1 (African green monkey kidney fibroblast-like cell line) whole cell lysates
Lane 4 : MDCK (Canine kidney cell line) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
-
![Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)](https://www.abcam.com/ps/products/181/ab181603/Images/ab181603-230547-anti-gapdh-antibody-epr16884-loading-control-western-blot.jpg)
Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)All lanes : Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603) at 1/50000 dilution
Lane 1 : UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates
Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
-
![Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)](https://www.abcam.com/ps/products/181/ab181603/Images/ab181603-230545-anti-gapdh-antibody-epr16884-loading-control-western-blot.jpg)
Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)All lanes : Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603) at 1/10000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Rat brain lysate
Lane 4 : Rat heart lysate
Lane 5 : Rat kidney lysate
Lane 6 : Rat spleen lysate
Lane 7 : C6 (Rat glial tumor cells) whole cell lysates
Lane 8 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
-
![Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)](https://www.abcam.com/ps/products/181/ab181603/Images/ab181603-230544-anti-gapdh-antibody-epr16884-loading-control-western-blot.jpg)
Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)All lanes : Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603) at 1/10000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
-
![Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)](https://www.abcam.com/ps/products/181/ab181603/Images/ab181603-230538-anti-gapdh-antibody-epr16884-loading-control-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling GAPDH with ab181603 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm and nuclear staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
1. ab181603 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)](https://www.abcam.com/ps/products/181/ab181603/Images/ab181603-230543-anti-gapdh-antibody-epr16884-loading-control-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)Immunohistochemical analysis of paraffin-embedded Human transitional cell carcinoma of bladder tissue labeling GAPDH with ab181603 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic and nucleus staining on the tumor cells of transitional cell carcinoma of Human bladder is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)](https://www.abcam.com/ps/products/181/ab181603/Images/ab181603-230542-anti-gapdh-antibody-epr16884-loading-control-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling GAPDH with ab181603 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocytes of mouse spleen is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)](https://www.abcam.com/ps/products/181/ab181603/Images/ab181603-230540-anti-gapdh-antibody-epr16884-loading-control-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling GAPDH with ab181603 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocyte of rat spleen is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)](https://www.abcam.com/ps/products/181/ab181603/Images/ab181603-230536-anti-gapdh-antibody-epr16884-loading-control-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling GAPDH with ab181603 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
-
![Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)](https://www.abcam.com/ps/products/181/ab181603/Images/ab181603-10-benefits-of-recombinant-antibodies.png)
Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (146)
ab181603 has been referenced in 146 publications.
- Tian L et al. Long non-coding RNA LINC01006 exhibits oncogenic properties in cervical cancer by functioning as a molecular sponge for microRNA-28-5p and increasing PAK2 expression. Int J Mol Med 47:N/A (2021). PubMed: 33576457
- Azizidoost SH et al. Impact of Methyl-ß-Cyclodextrin and Apolipoprotein A-I on The Expression of ATP-Binding Cassette Transporter A1 and Cholesterol Depletion in C57BL/6 Mice Astrocytes. Cell J 23:93-98 (2021). PubMed: 33650825
- Yu H et al. LINC00504 Promotes the Malignant Biological Behavior of Breast Cancer Cells by Upregulating HMGB3 via Decoying MicroRNA-876-3p. Cancer Manag Res 13:1803-1815 (2021). PubMed: 33654429
- Tian T et al. FAST1 Predicts Poor Survival of Renal Carcinoma and Promotes Its Progression Through the TGF-ß/Smad Pathway. Onco Targets Ther 14:1487-1499 (2021). PubMed: 33679133
- Mosqueira M et al. Cardiomyocyte depolarization triggers NOS-dependent NO transient after calcium release, reducing the subsequent calcium transient. Basic Res Cardiol 116:18 (2021). PubMed: 33728868