Anti-beta 2 Microglobulin antibody (ab6608)

ADDITIONAL NOTES I only would like to know which size i am suposed to detect in western blotting with this antibody. Is b2microglobulin coupled to 44kDa HLA chain to form a complex about 56kDa? Is 44kDa the size of the complex detected with this antibody? Thank you

ANSWER:

Thank you for yur enquiry and your interest in our products.

The antibody reacts with b2-microglobulin isolated from human urine showing a 12.8 kDa band under non-reducing condition of SDS-PAGE.

Thank you for your replacement AB6608 which our customer has received in the meantime. Unfortunately the problem (many spots on the blot) does also appear with this new vial (which is of the same lot 7396)! So the question is, what is the cause for this problem? Do you have any more suggestions or feedback from your lab/QC?

Kind regards,

ANSWER:

Thank you for your patience.

Actually we've never seen a blot like this before. We consulted with a couple of senior staffs in our company. We came out several suggestions as following:

1. Filter the whatever blocking reagent they use, may add 10% Goat serum to the blocking buffer, do the blocking for at least 1 h, R.T.

2. Do a 1st antibody negative control blot side-by-side with a complete WB

2. Since the specific bands are so strong, further dilute the antibody. The primary antibody incubation time can be shortened to 1h, R.T.

There are many factors beyond our control. Please let us know if these could help.

Thank you.

Antibody AB6608 (lot 7396) showed many small 'spots' in WB (ECL) - other antibodies used in comparable experiments worked fine. See attached complaint form for details.

ANSWER:

Thank you for your e-mail.

We have solved several vial form exactly the same batch number without any problem.

Has this customer performed an experiment using different primary antibodies on the same blot? Has he done a head-to-head experiment?

We regret to inform you that usually having spots on the blot due to some practical problems:

1. Contamination. Customer should check that all the buffers are free from bacterial contamination and infection. Deionized water should be coming from a well maintained supply. 2. Not enough of the solution during incubation or washing. Make sure that the membrane is well emerged in the solution which is moving gently across the membrane. 3. During incubation air bubbles became trapped on the surface of the blot. Next time, gently remove all air bubbles.

Should you need any more help then please do let us know.