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GAPDH Antibody

Catalog # NB300-320 | Novus Biologicals a Bio-Techne Brand
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NB300-320

Key Product Details

Species Reactivity

Human, Mouse, Rat, Canine, Drosophila, Insect

Applications

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Peptide ELISA, Single Cell Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Concentration

0.5 mg/ml

Product Summary for GAPDH Antibody

Immunogen

This GAPDG antibody was developed against the peptide sequence C-HQVVSSDFNSDT corresponding to C-Terminus according to NP_002037.2.

Predicted Species

Porcine (100%). Backed by our 100% Guarantee.

Reactivity Notes

Drosophilia reactivity reported in scientific literature (PMID: 28888970 and PMID: 33046910). Canine reactivity reported in scientific literature (PMID: 30030415). Use in Mouse reported in scientific literature (PMID:32771388).

Localization

Cytoplasmic

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Theoretical MW

36 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for GAPDH Antibody

Western Blot: GAPDH Antibody [NB300-320] - Staining of Human Liver (A) and Tonsil (B) with antibody at 0.001 ug/mL and Rat Brain lysate (C) with antibody at 0.3 ug/mL (35 ug protein in RIPA buffer). Detected by chemiluminescence. Approx. 37 kDa band observed in Rat Brain lysates and approx. 35 kDa in Human Liver and Tonsil lysates (calculated MW of 36.1 kDa according to Human NP_002037.2 and 35.8 kDa according to Rat NP_058704.1).
Immunocytochemistry/Immunofluorescence: GAPDH Antibody [NB300-320] - Immunofluorescence analysis of paraformaldehyde fixed HeLa cells, permeabilized with 0.15% Triton. Primary incubation 1hr (10 ug/mL) followed by Alexa Fluor 488 secondary antibody (2 ug/mL), showing cytoplasmic and vesicle staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10 ug/mL) followed by Alexa Fluor 488 secondary antibody (2 ug/mL). Strong expression of the protein seen in the cytoplasm and vesicles of HeLa cells.
Immunohistochemistry-Paraffin: GAPDH Antibody [NB300-320] - Staining of paraffin embedded Human Pancreas. Antibody at 2.5 ug/mL. Steamed antigen retrieval with citrate buffer pH 6, AP-staining.

Applications for GAPDH Antibody

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

10 ug/mL

Immunohistochemistry-Paraffin

2.5 ug/mL

Peptide ELISA

Detection limit 1:128000

Single Cell Western

100 ug/ml

Western Blot

0.001 - 0.003 ug/mL
Application Notes
Single Cell Western reported by an internal validation on HeLa-GFP cells at a concentration of 100 ug/ml
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 4.5 using NB300-320 in the following applications:

Published Applications

Read 64 publications using NB300-320 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris saline (20 mM Tris pH 7.3, 150 mM NaCl), 0.5% BSA

Preservative

0.02% Sodium Azide

Concentration

0.5 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: GAPDH

Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) is a ubiquitous enzyme involved in glycolysis, converting glyceraldehyde-3-phosphate into 1,3 diphosphoglycerate. This constitutively expressed, homotetramer protein can be found in the nucleus and cytoplasm, and the monomer has a theoretical molecular weight of 36 kDa. Known as a housekeeping gene, GAPDH routinely serves as a control for real-time PCR (RT-PCR) and as a loading control for Western Blots (1-3). In addition to its role in glycolysis, GAPDH has multiple cellular functions including membrane trafficking, transcription activation, apoptosis, DNA repair, and has been implicated in various pathologies such as cancer and neurodegenerative diseases including Alzheimer's and Huntington's (4,5).

References

1) Barber RD, Harmer DW, Coleman RA, Clark BJ. (2005) GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiol Genomics. 21(3):389-95. PMID: 15769908

2) Jia Y, Takimoto K. (2006) Mitogen-activated protein kinases control cardiac KChIP2 gene expression. Circ Res. 98(3):386-93. PMID: 16385079

3) Godsel LM, Hsieh SN, Amargo EV, Bass AE, Pascoe-McGillicuddy LT, Huen AC, Thorne ME, Gaudry CA, Park JK, Myung K, Goldman RD, Chew TL, Green KJ. (2005) Desmoplakin assembly dynamics in four dimensions: multiple phases differentially regulated by intermediate filaments and actin. J Cell Biol. 171(6):1045-59. PMID: 16365169

4) Sirover MA1. (1999) New insights into an old protein: the functional diversity of mammalian glyceraldehyde-3-phosphate dehydrogenase. Biochim Biophys Acta. 1432(2): 159-84. PMID: 10407139

5) Tristan C, Shahani N, Sedlak TW, Sawa A. (2011) The diverse functions of GAPDH: views from different subcellular compartments. Cell Signal. 23(2):317-23. PMID: 20727968

Long Name

Glyceraldehyde-3-phosphate Dehydrogenase

Alternate Names

G3PDH

Entrez Gene IDs

2597 (Human); 14433 (Mouse); 24383 (Rat)

Gene Symbol

GAPDH

Product Documents for GAPDH Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for GAPDH Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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