Cell Signaling Technology

Product Pathways - Antibody Sampler Kits

Loading Control Antibody Sampler Kit (HRP Conjugate) #4670

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
β-Actin (13E5) Rabbit mAb (HRP Conjugate) #5125 40 µl W H M R Mk B Pg (C) (Hr) 45 Rabbit IgG
COX IV (3E11) Rabbit mAb (HRP Conjugate) #5247 40 µl W H Mk 17 Rabbit IgG
Histone H3 (3H1) Rabbit mAb (HRP Conjugate) #5192 40 µl W H M R Hm Mk Z B Pg 17 Rabbit IgG
β-Tubulin (9F3) Rabbit mAb (HRP Conjugate) #5346 40 µl W H M R Mk Z B (C) 55 Rabbit IgG
GAPDH (D16H11) XP® Rabbit mAb (HRP Conjugate) #8884 40 µl W H M R Mk 37 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  C=Chicken  Z=Zebrafish  B=Bovine  Pg=Pig  Hr=Horse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each antibody in the Loading Control Antibody Sampler Kit (HRP Conjugate) detects endogenous levels of its target protein and does not typically cross-react with other proteins. β-Actin (13E5) Rabbit mAb (HRP Conjugate) detects endogenous levels of total β-actin protein and may cross-react with γ-actin.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using β-Actin (13E5) Rabbit mAb (HRP Conjugate) #5125.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H3 (3H1) Rabbit mAb (HRP Conjugate) #5192.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat, HeLa and COS cells using COX IV (3E11) Rabbit mAb (HRP Conjugate) #5247.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using β-Tubulin (9F3) Rabbit mAb (HRP Conjugate) #5346.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 and HeLa cells using GAPDH (D16H11) XP® Rabbit mAb (HRP Conjugate) #8884.

Description

The Loading Control Antibody Sampler Kit (HRP Conjugate) contains antibodies directed against a variety of housekeeping proteins. These Cell Signaling Technology antibodies are conjugated to the carbohydrate groups of horseradish peroxidase (HRP) via its amine groups. The kit contains enough primary antibodies to perform four western blots.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues near the amino terminus of human β-actin, surrounding Lys29 of human COX IV, near the carboxy terminus of human GAPDH, near the carboxy terminus of human histone H3, and near the amino terminus of human β-tubulin.

Background

Housekeeping proteins perform numerous basic functions within the cell and are constitutively expressed at high levels in a variety of tissues and cell types. Western blot analysis commonly uses housekeeping proteins such as β-actin, COX IV, GAPDH, histone H3 and the α- and β-tubulins as loading controls. Actin is a ubiquitous protein and a major component of the eukaryotic cytoskeleton. Actin exists mainly as the F-actin fibrous polymer (1). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyzes the phosphorylation of glyceraldehyde-3-phosphate during glycolysis. Recent work has demonstrated that GAPDH plays roles in apoptosis (2), gene expression (3), and nuclear transport (4). Globular tubulin subunits made up of α- and β-tubulin heterodimers are the building blocks of microtubules, one of three types of cytosolic fibers that comprise the cytoskeleton (5). Histone proteins, including histone H3, make up the primary building block of chromatin known as nucleosomes. Modulation of the chromatin structure plays an important role in the regulation of transcription in eukaryotes (6). Cytochrome c oxidase (COX) is a hetero-oligomeric enzyme consisting of 13 subunits localized to the inner mitochondrial membrane (7-9). It is the terminal enzyme complex in the respiratory chain, catalyzing the reduction of protons across the mitochondrial inner membrane to drive ATP synthesis (10).

  1. Condeelis, J. (2001) Trends Cell Biol 11, 288-93.
  2. Hara, M.R. and Snyder, S.H. Cell Mol Neurobiol 26, 527-38.
  3. Zheng, L. et al. (2003) Cell 114, 255-66.
  4. Bae, B.I. et al. (2006) Proc Natl Acad Sci U S A 103, 3405-9.
  5. Westermann, S. and Weber, K. (2003) Nat Rev Mol Cell Biol 4, 938-47.
  6. Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79.
  7. Ostermeier, C. et al. (1996) Curr Opin Struct Biol 6, 460-6.
  8. Capaldi, R.A. et al. (1983) Biochim Biophys Acta 726, 135-48.
  9. Kadenbach, B. et al. (2000) Free Radic Biol Med 29, 211-21.
  10. Barrientos, A. et al. (2002) Gene 286, 53-63.

Application References

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Protocols

Companion Products

Rabbit monoclonal antibodies are produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and 7,429,487) from Epitomics, Inc.

For Research Use Only. Not For Use In Diagnostic Procedures.

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