Anti-JAK2 antibody (ab39636)

ab39636 JAK antibody not working in IHC. Staining is weak and diffuse.

ANSWER:

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

I would like to reassure you that this antibody is tested and covered by our guarantee for IHC-P, IHC-Fr and and in Mouse, Rat, Dog and Human samples.In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

In this case, I woudl be pleased to provide afree of charge replacement or credit note in compensation if the antibody has been used in a tested species.

I would also appreciate if you are able to provide some further information which is beneificial for our quality monitoring records:

1. Which type of tissue has been tested?

2. What dilutions has the antibody been used at?

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

I have a question regarding the treatment of tissue prior to applying the primary antibody. Do you recommend microwave? Enzymatic?

The tissue I have in mind is human salivary gland.

I'm planning to use ab6672, ab59389, ab39636, ab32101, ab31370 and ab30646. Different recommendations?

ANSWER:

Thank you for contacting us.

The treatment of PFA-fixed, paraffin embedded sections (IHC-P) before incubation with priamry antibody would include antigen retrieval, blocking of endogenous peroxidases (if HRP is being used for detection), and blocking of non-specific proteins. Here are the protocol details for the respective antibodies:

ab30646, ab31370, ab39636, ab59389:

1. antigen retrieval: 10 mM citrate buffer (pH6.0) boil in pressure cooker

2. block with 3% H2O2 (in fresh methanol) for 15 minutes at room temperature

3. block with 3% BSA in TBS for 30 minutes

ab6672:

antigen retrieval: citrate buffer pH 6 and proteinase K digestion have been tried successfully as well as no antigen retrieval.

2. block with 3% hydrogen peroxide for 10 min

3. block with 1-10% Serum for up to 1 hour

ab32101:

1. antigen retrieval:10 mM Sodium Citrate Buffer, pH 6.0 in rice cooker

2. block with 3% hydrogen peroxide for 10 min

3. block with PBS + 10% serum

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.