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5142
Loading Control Antibody Sampler Kit

Loading Control Antibody Sampler Kit #5142

Western Blotting Image 1

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

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Western Blotting Image 2

Western blot analysis of extracts from COS-7, NIH/3T3 and PC12 cells, using β-Tubulin (9F3) Rabbit mAb.

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Western Blotting Image 3

Western blot analysis of extracts from various cell lines using Histone H3 (D1H2) XP® Rabbit mAb.

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Western Blotting Image 4

Western blot analysis of extracts from various cell lines using GAPDH (D16H11) XP® Rabbit mAb.

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Western Blotting Image 5

Western blot analysis of extracts from various cell lines using β-Actin (D6A8) Rabbit mAb.

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Western Blotting Image 6

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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IHC-P (paraffin) Image 7

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing staining of the mitochondria, using COX IV (3E11) Rabbit mAb.

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IHC-P (paraffin) Image 8

Immunohistochemical analysis of paraffin-embedded human glioblastoma using β-Tubulin (9F3) Rabbit mAb.

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IHC-P (paraffin) Image 9

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Histone H3 (D1H2) XP® Rabbit mAb.

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IHC-P (paraffin) Image 10

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using GAPDH (D16H11) XP® Rabbit mAb.

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Flow Cytometry Image 11

Flow cytometric analysis of NIH/3T3 cells using β-Actin (D6A8) Rabbit mAb (blue) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.

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IHC-P (paraffin) Image 12

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using COX IV (3E11) Rabbit mAb in the presence of control peptide (left) or Cox IV Blocking Peptide #1034 (right).

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IHC-P (paraffin) Image 13

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using β-Tubulin (9F3) Rabbit mAb.

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Flow Cytometry Image 14

Flow cytometric analysis of human peripheral blood lymphocytes using Histone H3 (D1H2) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.

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IHC-P (paraffin) Image 15

Immunohistochemical analysis of paraffin-embedded mouse colon using GAPDH (D16H11) XP® Rabbit mAb.

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IF-IC Image 16

Confocal immunofluorescent analysis of HeLa cells using β-Actin (D6A8) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IHC-P (paraffin) Image 17

Immunohistochemical analysis of paraffin-embedded H1650 xenograft, using COX IV Rabbit mAb. Note specific staining of human cancer cells.

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IHC-P (paraffin) Image 18

Immunohistochemical analysis of paraffin-embedded human melanoma using β-Tubulin (9F3) Rabbit mAb.

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IF-IC Image 19

Confocal immunofluorescent analysis of HeLa cells using Histone H3 (D1H2) XP® Rabbit mAb (green) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red).

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IF-IC Image 20

Confocal immunofluorescent analysis of C2C12 cells using GAPDH (D16H11) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IHC-F (frozen) Image 21

Immunohistochemical analysis of frozen H1650 xenograft, using Cox IV (3E11) Rabbit mAb.

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IHC-P (paraffin) Image 22

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using β-Tubulin (9F3) Rabbit mAb preincubated with control peptide (left) or β-Tubulin Blocking Peptide #1032 (right).

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Flow Cytometry Image 23

Flow cytometric analysis of HeLa cells, using COX IV (3E11) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

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Flow Cytometry Image 24

Flow cytometric analysis of NIH/3T3 cells using β-Tubulin (9F3) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

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IF-IC Image 25

Confocal immunofluorescent analysis of HeLa cells labeled with COX IV (3E11) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IF-IC Image 26

Confocal immunofluorescent analysis of HeLa cells using β-Tubulin (9F3) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
COX IV (3E11) Rabbit mAb 4850 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H R Mk Z B Pg 17 Rabbit IgG
β-Tubulin (9F3) Rabbit mAb 2128 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Mk Z B 55 Rabbit IgG
Histone H3 (D1H2) XP® Rabbit mAb 4499 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Mk 17 Rabbit IgG
GAPDH (D16H11) XP® Rabbit mAb 5174 20 µl
  • WB
  • IHC
  • IF
H M R Mk 37 Rabbit IgG
β-Actin (D6A8) Rabbit mAb 8457 20 µl
  • WB
  • IF
  • F
H M R Mk Dm Z 45 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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

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.

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, and 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).

  1. Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79.
  2. Condeelis, J. (2001) Trends Cell Biol. 11, 288-293.
  3. Hara, M.R. and Snyder, S.H. (2006) Cell Mol. Neurobiol. 26, 527-38.
  4. Westermann, S. and Weber, K. (2003) Nat Rev Mol Cell Biol 4, 938-47.
  5. Capaldi, R.A. et al. (1983) Biochim. Biophys. Acta 726, 135-148.
  6. Zheng, L. et al. (2003) Cell 114, 255-66.
  7. Ostermeier, C. et al. (1996) Curr. Opin. Struct. Biol. 6, 460-466.
  8. Kadenbach, B. et al. (2000) Free Radic. Biol. Med. 29, 211-221.
  9. Bae, B.I. et al. (2006) Proc. Natl. Acad. Sci. USA 103, 3405-9.
  10. Barrientos, A. et al. (2002) Gene 286, 53-63.
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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