Anti-GAPDH Antibody (HRP)

Product Name

Anti-GAPDH Antibody (HRP)

View all GAPDH Antibodies

SKU/Catalog Number

A00227-HRP

Size

100μg/vial

Form

Liquid

Description

Boster Bio Anti-GAPDH Antibody (HRP) (Catalog # A00227-HRP). Tested in WB applications. This antibody reacts with Human, Mouse, Rat, Monkey, Chicken, Zebrafish.

Storage & Handling

HRP-GAPDH antibody can be stored at 4°C for three months and -20°C, stable for up to one year.

Cite This Product

Anti-GAPDH Antibody (HRP) (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00227-HRP)

Host

Rabbit

Contents

Each vial contains 50% glycerol, 0.9% NaCl, 0.2% Na2HPO4.

Clonality

Polyclonal

Clone Number

WH-3

Isotype

IgG

Immunogen

HRP-GAPDH antibody was raised against a synthetic peptide containing 16 amino acids near the carboxy terminus of GAPDH.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

HRP-GAPDH antibody is human, mouse, rat, rabbit, and chicken reactive.

Reactive Species

A00227-HRP is reactive to GAPDH in Chicken, Human, Monkey, Mouse, Rat, Zebrafish

Applications

A00227-HRP is guaranteed for WB Boster Guarantee

Observed Molecular Weight

36 kDa

Calculated molecular weight

36.053kDa

Background of GAPDH

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyzes the reversible oxidative phosphorylation of glyceraldehyde-3-phosphate in the presence of inorganic phosphate and nicotinamide adenine dinucleotide (NAD), an important energy-yielding step in carbohydrate metabolism. It also is involved in a number of cellular processes such as membrane fusion, phosphotransferase activity, DNA replication and repair, and nuclear RNA export (1). GAPDH also plays a role in different pathologies such as cancer progression, apoptosis, and neuronal diseases such as Alzheimer's and Huntington's disease (2). GAPDH is constitutively expressed at high levels in almost all tissues and cell lines making it ideal for use as a loading control marker in immunoblots.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.02-0.1 μg/ml

Validation Images & Assay Conditions

Click image to see more details

Figure 1. Western blot analysis of GAPDH using anti-GAPDH antibody (A00227-HRP).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human A549 whole cell lysates,
Lane 3: human A562 whole cell lysates,
Lane 4: human MCF-7 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat liver tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse liver tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GAPDH antigen affinity purified polyclonal antibody (Catalog # A00227-HRP) at 0.1 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for GAPDH at approximately 36 kDa. The expected band size for GAPDH is at 36 kDa.

Click image to see more details

Figure 2. Western blot analysis of GAPDH using anti-GAPDH antibody (A00227-HRP).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: chicken heart tissue lysates,
Lane 2: chicken liver tissue lysates,
Lane 3: chicken brain tissue lysates,
Lane 4: monkey heart tissue lysates,
Lane 5: monkey lung tissue lysates,
Lane 6: monkey kidney tissue lysates,
Lane 7: monkey liver tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GAPDH antigen affinity purified polyclonal antibody (Catalog # A00227-HRP) at 0.1 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for GAPDH at approximately 36 kDa. The expected band size for GAPDH is at 36 kDa.

Gene/Protein Information For GAPDH (Source: Uniprot.org, NCBI)

Alternative Names

1D4 GAPDH; aging-associated gene 9 protein; Cytoplasm Marker; EC 1.2.1; EC 1.2.1.12; EC 2.6.99.-; G3PD; G3PD; G3PDH; GAPD; GAPDH; GAPDPeptidyl-cysteine S-nitrosylase GAPDH; glyceraldehyde 3-phosphate dehydrogenase; glyceraldehyde-3-phosphate dehydrogenase; MGC102544; MGC102546; MGC103190; MGC103191; MGC105239; MGC127711; MGC88685; OCAS, p38; OCT1 coactivator in S phase; Peptidyl-cysteine S-nitrosylase GAPDH GAPDH G3PD, GAPD, HEL-S-162eP glyceraldehyde-3-phosphate dehydrogenase glyceraldehyde-3-phosphate dehydrogenase|OCAS, p38 component|Oct1 coactivator in S phase, 38 Kd component|aging-associated gene 9 protein|epididymis secretory sperm binding protein Li 162eP|peptidyl-cysteine S-nitrosylase GAPDH

For more info on GAPDH, check out the GAPDH Infographic

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In this infographic, you will see the following information for GAPDH: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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