Name: Anti-GAPDH antibody [EPR16884] - Loading Control
Brand: RabMAb
SKU: ab181603
Rating: 5 (6 reviews)

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: WB, IHC-P, ICC/IF, Flow Cytmore details
Species reactivity

Reacts with: Mouse, Rat, Chicken, Cow, Dog, Human, Monkey
Immunogen

Recombinant fragment within Mouse GAPDH aa 1-150. The exact sequence is proprietary.
Database link: P16858

Run BLAST with Run BLAST with
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

Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab^® patents.
This product is a recombinant rabbit monoclonal antibody.

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

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
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.
Flow Cyt		1/200. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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 accession P04406 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

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
- Omim: 138400 Human
- SwissProt: P00356 Chicken
- SwissProt: P10096 Cow
- SwissProt: Q28259 Dog
- SwissProt: P04406 Human
- SwissProt: P16858 Mouse
- SwissProt: P04797 Rat
- Unigene: 544577 Human
- Unigene: 592355 Human
- Unigene: 598320 Human
- Unigene: 304088 Mouse
- Unigene: 309092 Mouse
- Unigene: 317779 Mouse
- Unigene: 343110 Mouse
- Unigene: 392463 Mouse
- Unigene: 392480 Mouse
- Unigene: 414470 Mouse
- Unigene: 458138 Mouse
- Unigene: 458416 Mouse
- Unigene: 475698 Mouse
- Unigene: 129558 Rat
- Unigene: 91450 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
- 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

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 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.
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 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.
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 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.
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 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.
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)

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.
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.
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.
Flow Cytometry - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)

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.

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

This product has been referenced in:

Lv L et al. MicroRNA-495 targets Notch1 to prohibit cell proliferation and invasion in oral squamous cell carcinoma. Mol Med Rep 19:693-702 (2019). Read more (PubMed: 30387817) »
Cai W et al. MicroR-542-3p can mediate ILK and further inhibit cell proliferation, migration and invasion in osteosarcoma cells. Aging (Albany NY) 11:18-32 (2019). Read more (PubMed: 30636169) »

See all 29 Publications for this product

Publishing research using ab181603? Please let us know so that we can cite the reference in this datasheet.

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1-6 of 6 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Immunocytochemistry/ Immunofluorescence

Sample

Mouse Cell (undifferentiated Human ES cells on MEFs)

Permeabilization

Yes - see blocking buffer

Specification

undifferentiated Human ES cells on MEFs

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 14 2016

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Human Cell lysate - whole cell (HUES7 ES cells)

Gel Running Conditions

Reduced Denaturing (12%)

Loading amount

20 µg

Specification

HUES7 ES cells

Blocking step

Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C

Abcam user community

Verified customer

Submitted Feb 04 2016

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (HEPG2)

Specification

HEPG2

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 18 2015

Application

Western blot

Sample

Human Cell lysate - whole cell (HEPG2)

Gel Running Conditions

Reduced Denaturing (12.5)

Loading amount

20 µg

Specification

HEPG2

Blocking step

Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C

Abcam user community

Verified customer

Submitted Nov 13 2015

Application

Immunocytochemistry/ Immunofluorescence

Blocking step

Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 16°C

Sample

Mouse Cell (mouse embryonic stem cell)

Specification

mouse embryonic stem cell

Permeabilization

Yes - 0.1% triton X

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 18 2014

Application

Western blot

Loading amount

100000 cells

Gel Running Conditions

Reduced Denaturing (10% gel)

Sample

Mouse Cell lysate - whole cell (mouse embryonic stem cell)

Specification

mouse embryonic stem cell

Blocking step

Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C

Abcam user community

Verified customer

Submitted Aug 08 2014

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

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Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)

Overview

Properties

Associated products

Applications

Target

Images

Protocols

Datasheets and documents

References

This product has been referenced in:

Customer reviews and Q&As

Immunocytochemistry/ Immunofluorescence abreview for Anti-GAPDH antibody [EPR16884]

Western blot abreview for Anti-GAPDH antibody [EPR16884]

Immunocytochemistry/ Immunofluorescence abreview for Anti-GAPDH antibody [EPR16884]

Western blot abreview for Anti-GAPDH antibody [EPR16884]

Immunocytochemistry/ Immunofluorescence abreview for Anti-GAPDH antibody [EPR16884]

Western blot abreview for Anti-GAPDH antibody [EPR16884]