Anti-Chemokine Receptor D6抗体(ab1656)

Thank you for submitting an Abreview of ab1656. As you may have noticed, your review has now been published on our website.
Since you obtained poor results using the antibody in a untested species, we would like to follow up on this to see if we can possibly improve the results by offering some troubleshotting advice.
Based on the information in your review, I feel that altering the following step in your protocol may help improve your results:
- the homology of the immunogen with rat is 89%, thus, theoretically might work, but dependend on where exactly the epitope the binding might be weak.
- check if the protein is expressed with high enough copy numbers in this sample type
- incubate longer with the primary antibody
I thank you for your Abreview submission and wish you good luck with your research.

Thank you for calling Abcam yesterday.
I have talked to the lab and I am sorry but we do not have an immunohistochemistry image showing the use of the blocking peptide with the antibody.
As we talked about yesterday as well, please find below the links that explain about our Abpromise and our testing discount program:
https://www.abcam.com/index.html?pageconfig=abpromise
https://www.abcam.com/index.htmlpageconfig=resource&rid=11998&viapagetrap=collaborationdiscount
If there is anything else I can help you with, or if you would like to take advantage of our testing discount program to try ab1656 in western blot, please let me know.

Thank you for your email and I'm sorry to hear that you are still experiencing difficulty with ab1656. You should definitely see positive staining with the spleen and lung tissue and so it seems as if the antibody is damaged in some way. I can have a refund issued to you - can you please give me the purchase order number that was used when you ordered this antibody? Thank you and I look forward to hearing from you.

Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab1656. Our source for this antibody was able to provide me with the IHC-P protocol below, which I hope will be helpful. Also, mouse spleen or lung tissue is recommended as a positive control. If you need additional assistance, please contact us again. Immunohistochemistry on Paraffin sections: Paraffin sections of tissues fixed in 10% formalin were used for testing antibodies. Preparation of slides: Place slides in 60 C oven overnight after cutting. Then follow the procedure below: 1) xylene 2x10min 2) 100% alcohol 2x10min 3) 95% alcohol 5 min 4) 80% alcohol 5 min 5) rinse in distilled water 2x2 min 6) antigen retrieval if necessary 7) Immunohistochemistry Antigen Retrieval: Sections were treated with antigen retrieval solution (Dako # S-1700, pH 6.0 or #S-3308, pH 8.0) and steamed for 20 minutes followed by cooling for 20 minutes. Immunohistochemistry: Block the endogenous peroxide by incubating the sections with 3% H2O2 for 15 minutes at room temperature (RT). Wash the sections with PBS and incubate with 2% normal rabbit serum at room temperature to block the non-specific binding sites. Wash the section with PBS and incubate with primary antibody (anti-chemokine receptor) at recommended dilution for 60 minutes at RT. Wash the slides and incubate at RT with biotinylated rabbit anti-goat IgG H+L (Vector Labs) at 1:200.

Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab1656. Our source for this antibody was able to provide me with the IHC-P protocol below, which I hope will be helpful. Also, mouse spleen or lung tissue is recommended as a positive control. If you need additional assistance, please contact us again. Immunohistochemistry on Paraffin sections: Paraffin sections of tissues fixed in 10% formalin were used for testing antibodies. Preparation of slides: Place slides in 60 C oven overnight after cutting. Then follow the procedure below: 1) xylene 2x10min 2) 100% alcohol 2x10min 3) 95% alcohol 5 min 4) 80% alcohol 5 min 5) rinse in distilled water 2x2 min 6) antigen retrieval if necessary 7) Immunohistochemistry Antigen Retrieval: Sections were treated with antigen retrieval solution (Dako # S-1700, pH 6.0 or #S-3308, pH 8.0) and steamed for 20 minutes followed by cooling for 20 minutes. Immunohistochemistry: Block the endogenous peroxide by incubating the sections with 3% H2O2 for 15 minutes at room temperature (RT). Wash the sections with PBS and incubate with 2% normal rabbit serum at room temperature to block the non-specific binding sites. Wash the section with PBS and incubate with primary antibody (anti-chemokine receptor) at recommended dilution for 60 minutes at RT. Wash the slides and incubate at RT with biotinylated rabbit anti-goat IgG H+L (Vector Labs) at 1:200.