Hi Folks,
Could you please suggest me which is the best house keeping protein for Laccaria for western blot assay?
Does anybody has any antibody I may request or any suggestions please.....
Thank you in advance
Torikul Islam
Postdoc, Oakridge national Lab
USA
The best housekeeping protein for Laccaria bicolor fungi is a subjective matter and may vary depending on the experiment's aim and the research conditions. However, commonly used housekeeping proteins as internal controls in fungal studies include glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin, and beta-tubulin. These proteins are considered to be stable and expressed at relatively constant levels in most fungal cells, making them suitable as internal controls.
It's important to note that the expression levels of these housekeeping proteins may be affected by different factors, such as fungal growth stage, stress conditions, and cultural conditions, so it's essential to validate the suitability of a specific housekeeping protein for a given experiment. Additionally, it's advisable to use multiple housekeeping proteins to ensure robust normalization and minimize the effects of variability.
Hi Kunal, I really appreciate your answer. I have a quick question, so these house keeping proteins are commercially available? I mean, these actin, beta-tubulin are dedicated to fungi or we can use the antibodies used for plant as well??
Thank you again....
Yes, commercial antibodies against beta-actin, GAPDH, actin, and beta-tubulin are widely available. These antibodies are typically raised against mammalian or plant protein sequences and may cross-react with fungal homologs. It's essential to verify the specificity of the antibody for the target organism, especially when using antibodies raised against mammalian or plant protein sequences, as cross-reactivity can occur.
One approach to confirm antibody specificity is to perform a control Western blot using a sample from the target organism, to ensure that the antibody specifically recognizes the target protein and doesn't react with other proteins. Another approach is to perform a knockdown experiment, where the expression of the housekeeping protein is reduced, to demonstrate that the observed changes in protein levels are specific and not due to cross-reactivity with other proteins
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology
- Antibodies