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Loading Control Antibody Sampler Kit #5142

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
β-Actin (13E5) Rabbit mAb # 4970 40 microliters W IHC-P IHC-F IF-IC F H M R Mk Pg B (C) (Dg) (Hr) 45 Rabbit IgG
COX IV (3E11) Rabbit mAb # 4850 40 microliters W IP IHC-P IHC-F IF-IC F H R Mk Pg B Z 17 Rabbit IgG
GAPDH (14C10) Rabbit mAb # 2118 40 microliters W IHC-P IF-IC F H M R Mk 37 Rabbit IgG
Histone H3 (3H1) Rabbit mAb # 9717 40 microliters W H M R Mk Pg Hm B Z 17 Rabbit IgG
α-Tubulin (11H10) Rabbit mAb # 2125 40 microliters W IHC-P IF-IC F H M R Mk B Dm (Dg) 52 Rabbit IgG
β-Tubulin (9F3) Rabbit mAb # 2128 40 microliters W IHC-P IF-IC F H M R Mk B Dm Z (C) 55 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody # 7074 100 microliters Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Pg=Pig  C=Chicken  Hm=Hamster  B=Bovine  Dm=D. melanogaster  Z=Zebrafish  Dg=Dog  Hr=Horse

Specificity / Sensitivity

Each antibody in the Loading Control Antibody Sampler Kit detects endogenous levels of its target protein and does not typically cross-react with other proteins. β-Actin (13E5) Rabbit mAb 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 GAPDH (14C10) Rabbit mAb #2118.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using α-Tubulin (11H10) Rabbit mAb #2125.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS-7, NIH/3T3 and PC-12 cells using β-Tubulin (9F3) Rabbit mAb #2128.


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, Jurkat and COS cell lines using COX IV (3E11) Rabbit mAb #4850.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using β-Actin (13E5) Rabbit mAb #4970.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H3 (3H1) Rabbit mAb #9717.


Description

The Loading Control Antibody Sampler Kit contains antibodies to a variety of housekeeping proteins. The kit contains enough primary and secondary antibodies to perform four western blots per primary antibody.

Source / Purification

Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to residues near the amino terminus of human β-actin, surrounding Lys29 of human COX IV, the sequence of human GAPDH, the carboxy-terminal residues of human histone H3, the amino terminus of human α-tubulin and 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 (2006) 26, 527-38.
  3. Zheng, L. et al. (2003) Cell 114, 255-66.
  4. Bae, B.I. et al. (2006) Proc Natl Acad Sci USA 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 U.S.S.N. 11/476,277) from Epitomics, Inc.

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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