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Western Blot Loading Control Sampler Kit |
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| Catalog No: |
IMG-6166K |
| Contents: |
Primay antibodies contain 50 ug in 100 ul PBS with 0.05% BSA and 0.05% sodium azide. Secondary antibody contains 50 ul antibody in PBS and 0.05% BSA, no azide. BSA is of U.S. origin. |
| Isotype: |
Rabbit IgG |
| Clone: |
N/A |
| Purification: |
Protein G Chromatography |
| Species: |
Multi |
| Host: |
Rabbit |
| Application: |
Western blot analysis: Please see individual data sheets for details. |
| Storage: |
Store at 4°C, stable for 6 months. For long-term storage, store at -20°C. |
Recommended
Positive Control: |
Please see individual data sheets for details. |
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Background
Genes that are constitutively and stably expressed at high levels in almost all tissues and cells are often referred to as housekeeping genes. Housekeeping proteins such as GAPDH, beta Actin, beta Tubulin, or Histone H2B (nuclear fractions) are useful as loading controls for western blots and protein normalization. Increasingly, researchers are making use of specific antibodies to housekeeping proteins to standardize and compare levels of protein. However, studies examining gene transcript and protein levels have shown that expression levels of housekeeping genes can vary. Collectively the results indicate that some housekeeping proteins may be better suited than others for use as loading controls depending on the model system used.
The Western Blot Loading Control Sampler Kit is a cost-effective tool containing a panel of four antibodies specific to proteins frequently used for protein normalization, plus appropriate secondary reagent. Sufficient quantity of each antibody is included for several mini-blots; see individual datasheets for additional information on species reactivity and suggested dilutions.
| Description |
Cat. No. |
Qty |
Appl |
Reactivity |
Format |
| beta Actin |
IMG-5142A |
50 ug |
WB |
H, M, R |
Purified |
| beta Tubulin |
IMG-5810A |
50 ug |
WB |
Bo, Ch, D, H, M, Mk, R, Xe |
Purified |
| GAPDH |
IMG-5143A |
50 ug |
WB |
Ca, Chp, Dros, H, M, Mq, Pi, R |
Purified |
| Histone H2B |
IMG-359 |
50 ug |
WB |
Ch, D, Dros, H, M, Ma, R, Zf |
Purified |
| Anti-Rabbit Ig HRP Secondary |
20301 |
50 ul |
WB |
Ra |
Purified |
| Bo: Bovine Ca:Cat Ch:Chicken Chp:Chimpanzee D:Dog Dros:Drosophila H:Human M:Mouse Ma:Mammals Mk:Monkey Mq:Macaque R:Rat Xe:Xenopus Zf:Zebrafish | |
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Western blot analysis of beta actin in A: human brain (40141), B: mouse brain (40121), C: rat brain (40121), D: human lung (40146) and E: human spleen (40149) tissue lysates using IMG-5142A at 0.25 ug/ml. |
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Western blot analysis of beta tubulin in A: human brain (40141), B: mouse brain (40121) and C: rat brain (40121) tissue lysates using IMG-5810A at 0.25 ug/ml. |
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Western blot analysis of GAPDH in the multiple human tumor cell line lysate INSTA-Blot (IMB-105) using IMG-5143A at 0.25 ug/ml. |
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Reference
1. Housekeeping genes as internal standards: Use and limits. Thellin O, W Zorzi, B Lakaye, B De Borman, B Coumans, G Hennen, T Grisar, A Igout, and E Heinen. J Biotechnology 75:291-295 (1999).
2. Housekeeping proteins: A preliminary study illustrating some limitations as useful references in protein expression studies. Ferguson RE, HP Carroll, A Harris, EM Maher, PJ Selby, and RE Banks. Proteomics 5:566-571 (2005).
3. Western blotting. Kurien, BT and RH Scofield. Methods 38:283-293 (2006).
4. GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Barber RD, DW Harmer, RA Coleman and BJ Clark. Physiol Genomics 21:389-395 (2005).
5. An error model for protein quantification. Kreutz C, MM Bartolome Rodriquez, T Maiwald, M Seidi, HE Blum, L Mohr, and J Timmer. Bioinformatics 23:2747-2753 (2007).
6. De-regulation of common housekeeping genes in heptocellular carcinoma. Waxman S and E Wurmbach. BMC Genomics doi:10.1186/1471-2164-8-243 (2007).
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Research purposes only.
Not for diagnostic or in vivo use.
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