Western blot analysis of extracts from Jurkat, HeLa and COS cell lines, using COX IV (3E11) Rabbit mAb (HRP Conjugate).
Western blot analysis of extracts from various cell lines using β-Tubulin (9F3) Rabbit mAb (HRP Conjugate).
Western blot analysis of extracts from NIH/3T3 and HeLa cells using GAPDH (D16H11) XP® Rabbit mAb (HRP Conjugate).
Western blot analysis of extracts from various cell lines using β-Actin (D6A8) Rabbit mAb (HRP Conjugate).
Western blot analysis of extracts from various cell lines using Histone H3 (D1H2) XP® Rabbit mAb (HRP Conjugate).
|COX IV (3E11) Rabbit mAb (HRP Conjugate) 5247||20 µl||
||H R Mk Z B Pg||17||Rabbit IgG|
|β-Tubulin (9F3) Rabbit mAb (HRP Conjugate) 5346||20 µl||
||H M R Mk Z B||55||Rabbit IgG|
|GAPDH (D16H11) XP® Rabbit mAb (HRP Conjugate) 8884||20 µl||
||H M R Mk||37||Rabbit IgG|
|β-Actin (D6A8) Rabbit mAb (HRP Conjugate) 12620||20 µl||
||H M R Mk Dm Z||45||Rabbit IgG|
|Histone H3 (D1H2) XP® Rabbit mAb (HRP Conjugate) 12648||20 µl||
||H M R Mk||17||Rabbit IgG|
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 two western blots.
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. Histone H3 (D1H2) XP® Rabbit mAb (HRP Conjugate) detects endogenous levels of total histone H3 protein, including the Histone H3 variant CENP-A.
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.
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).
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