Anti-CD90 / Thy1 antibody [F15-42-1] (FITC) (ab11155)

May I know if the following antibodies can be used for ICC and/or western blot? 1) CD31 (ab27333) 2) CD71 (ab47095) 3) CD44 (ab30437) 4) CD90 (ab11155) Also, can the IgG1(ab106163) be used as the isotype control for FACS with the 4 antibodies listed above? Many thanks in advance!

ANSWER:

Thank you very much for your interest in abcam antibodies. To our knowledge,these primary antibodies (ab27333, ab47095, ab30437 and ab11155)have not been tested in Western Blot and ICC/IF. They have been tested only in Flow cytometry as shown on the datasheets. Therefore, I can offer a discount off a future purchase if you buyone or allof themnow, test it/them in Western Blot and/or ICC/IF and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discountwould be to the value of 1 free primary antibody for each tested antibody. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and teston of those antibodiesin Western Blot and/or ICC/IF. I will then send a discount code. This code must be issued before purchasingthe antibodyso please wait for my reply before ordering. 2. Purchasethe primary antibody to be testedeither by phone, fax, or online (www.abcam.com). 3. Test it in Western Blot and/or ICC/IF. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: http://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even ifthe tested antibodyturns out to be unsuitable for Western Blotor ICC/IF, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount. I would like to confirm that ab106163 (Mouse IgG1 conjugated to FITC)can be used asisotype control with ab27333, ab47095, ab30437 and ab11155.

Sorry for forgetting to ask another question. I noted that the application of some antibodies is only for flow cytometry.CouldI use these antibodies to do western blot orICC? Thank you!

ANSWER:

If the antibody datasheets only list "flow cytometry" in the Applications section, then that is the only application that has been tested. It is possible that the antibodies could also work for western blotting or ICC, but these applications have not been tested, and they are not covered by our guarantee.

Thanks so much for your kind help. As I known, for flow cytometry, people always use normal cell(without antibody disposal) as control to set up the machine before running. But there are also some people use isotype antibody to replace normal cell as control. Which method is recommended for my control group to set up program? Many thanks!

ANSWER:

A cells-only control with no antibody will show the contribution of autofluorescence of the cells themselves. Most people use this to set up the flow cytometer. An isotype control (cells incubated with the same isotype of antibody as the target-specific antibody) will include autofluorescence signal but also the non-epitope-specific interaction of the antibody with the samples. Examples of these interactions include the binding of the Fc region of the antibody to Fc receptors on the cells, and any non-epitope-specific antibody-protein interactions (for instance, hydrophobic interactions) . You could use this isotype control to adjust your flow cytomer settings but most people include it as an experimental control, after setting up the flow cytometer with cells-only. You will be able to see if there is any signal from the isotype interactions by comparing the isotype control to the cells-only control. The antibodies conjugated to FITC that you asked about are all isotype IgG1 (ab30437, ab27333, ab11155). An appropriate isotype control is ab106163: Click here (or use the following: http://www.abcam.com/index.html?datasheet=106163). I hope this helps. If you have any questions, please contact me.

since these antibodies I choose are FITC-conjugated, need I use F4/1(ab112511) as anti-FITC antibody for flowcytomety set? Hope to hear from you soon. Many thanks!

ANSWER:

Thank you for contacting Abcam. I am replying to your question on behalf of my colleague Ada. The antibodies that have FITC conjugated to them do not need the anti-FITC antibody ab112511, if you simply want to detect FITC fluorescence by flow cytometry. If you have any questions or concerns, or if I have misunderstood your question, please reply.

                    Hi,   Our customer would like to know if the following antibodies are of IgG1, kappa  or lambda light chains (Information on datasheet only stated Isotypes IgG1/ IgM/ IgG2a)?   Item                                      Format                 Cat # Anti-CD200 antibody      unconjugated    ab17225 Anti-CD146 antibody      PE                           ab78488 Anti-CD34 antibody         unconjugated    ab6330 Anti-CD71 antibody         unconjugated    ab117720 Anti-CD24 antibody         FITC                       ab30350 Anti-CD73 antibody         unconjugated    ab81720                                                 PE                           ab106697 Anti-CD105 antibody      PE                           ab91238                                                 Unconjugated   ab11414 Anti-Cd45 antibody         PE                           ab90960 Anti-STRO1 antibody      unconjugated    ab102969 Anti-CD90 antibody         unconjugated    ab23894                                                 FITC                       ab11155 Anti-CD14 antibody         FITC                       ab86896               Anti-cytokeratin 15         unconjugated    ab1385 IgG2a Isotype control     FITC                       ab81197   Looking forward to your soonest reply. Thank you.   Best regards, Penny Hor BITA LIFESCIENCE SDN. BHD. C-23-05, Dataran 32, No. 2, Jalan 19/1, 46300 Petaling Jaya, Selangor Darul Ehsan, Malaysia t +603 7960 8286    e info@bitalifesc.com.my f +603 7960 8262     

ANSWER:

Thank you for contacting us.

In general, the immunoglobulin class (IgG1, IgG2a, IgGM, etc.) and light chain sub-class (kappa or lambda) is listed on the online datasheet. When the light chain is not listed, then the light chain sub-class has not been determined. In these instances, it is probably that the light chain sub-class is kappa, given that murine serum immunoglobulins are 95% kappa and 5% lambda, as discussed in the reference at the following link:

http://www.ncbi.nlm.nih.gov/pubmed/3116408

Mol Immunol. 1987 Jul;24(7):751-7. PMID:3116408

Of the antibodies your customer has enquired about, The light chain sub-class has been determined for three: ab90960, ab86896, and ab1385. All are kappa light chain.

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