Anti-Hsp70 antibody [3A3] (ab5439)

Thank you for your enquiry.

I would like to reassure you that both these antibodies are covered by our 6 month guarantee for WB and for human samples.

For more details regarding our guarantee, please see the following page from our website:

https://www.abcam.com/index.html?pageconfig=abpromise

As far as we are aware, the antibodies have not been specifically tried in WB with MCF7 cells. However, this is still likely to work, as they have been tested in WB and human, and so this would still be covered by our guarantee.

Also, you can obtain rewards by submitting an Abreview about these product via the online product datasheets. We always encourage customers to send their results back to us, whether positive or negative, and we make all product information available to researchers.

To find out more about our Abreview system and the Abpoints rewards you can obtain, please see the following webpage:

https://www.abcam.com/abreviews

I hope this will be helpful. Thank you for your time and effort.

Thank you for attending our webinar on Immunoprecipitation last week. We hope you enjoyed it and it was useful for you. We would also like to thank you for sending your question and apologize that we were not able to answer it in the Q&A session. The determination of the protein concentration before IP is not different from that for other techniques such as Western blot. Therefore, you could perform a Bradford assay, a Lowry assay or a BCA assay. Bovine serum albumin (BSA) is a frequently-used protein standard. Once you have determined the concentration of each sample, you can freeze them at -20°C or -80°C for later use or prepare for immunoprecipitation or for loading onto a gel. For the actual IP experiment, in a tube add 10-500 μg cell lysate plus the recommended amount of antibody. These amounts will be chosen depending on the abundance of the protein and the affinity of the antibody for the protein, typically in a pilot experiment where a fixed amount of protein is precipitated by increasing amounts of antibody. You can check the antibody datasheet for recommended antibody concentration. As a guideline we recommend: 1 – 5 μl polyclonal antiserum 1 μg affinity-purified polyclonal antibody 0.2 to 1 μl ascites fluid (monoclonal antibody) 20 to 100 μl culture supernatant (monoclonal antibody) I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for your message and for confirming these details. I am very pleased to hear the customer would like to accept our offer and test these antibodies in hamster. ab41684 Discount code: #### ab38975 Discount code: #### ab6314 Discount code: #### ab77179 Discount code: #### Expirey date for all these codes: 7th March 2012 This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit the Abreviews for hamster samples and include the relevant code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount. If you have any further questions, please do not hesitate to contact us. We look forward to receiving the Abreviews and wish the customer luck with their research.

Thank you for your message which has been forwarded to me as Karin is currently away from the office. I have reviewed these three antibodies with respect to discount codes: ab77179 I am sorry I am also unable to check the alignment of the immunogen with the hamster sequence, because there is regrettably no hamster sequence available on the protein database sites. I would be pleased to make an exception and provide a discount code in this case anyway, to test in Hamster. ab6314 There is 73% Alignment of the human full length immunogen to the hamster sequence you have provided. This is a little low, but there may be some crossreactivity so I would be pleased to provide a discount. ab16045 There is 14% alignment of the hamster sequence provided with the immunogen. This is very low so the antibody will probably not detect in hamster, and so I am sorry we would be unable to provide a discount for this particular antibody to test in hamster. I hope this will be helpful to you. Please confirm if the customer would like to receive a code for testing ab6314 and ab77179 in hamster and I will be pleased to arrange this for you.

Thank you for contacting us. If an antibody has not been tested with a particular species then a testing discount would apply. However, in order to check the likelihood of the antibody being reactive against the target the hamster protein sequence needs to be compared to that of the immunogen used to make sure that the most suitable antibody is chosen. Unfortunately I have been unable to find sequences for Entactin, Cathepsin L or Cyclophilin B for hamster so if you or your customer could help me with this that would be very useful. We already have several antibodies directed against Hsp70 which have been tested with hamster samples. ab47454 is a mouse monoclonal or ab79852 is a rabbit polyclonal. We also have several which are directly conjugated that may be of interest too. These can be found from the following link: https://www.abcam.com/index.html?pageconfig=searchresults&search=hsp70&pt=1&sk=app&sv=69&sn=WB&l=3&fViewMore=1 The Cyclophilin A antibody suggested (ab41684) is predicted to react and would therefore be suitable for a testing discount. However, with the TIMP2 antibody suggested (ab109708), the immunogen recognises the N-terminal sequence of the protein which differs significantly from that of the hamster protein. I would therefore recommend ab38975 as an alternative. This is a rabbit polyclonal directed against the C-terminal end which shares significant homology with Chinese hamster sequence (SwissProt Q60453) compared to the human (SwissProt P16035). I hope this information is helpful to you. If you'd like me to issue testing discounts for ab41684 or ab38975. Or you would like to stick with the original Hsp70 antibody recommended please let me know. Please do not hesitate to contact us if you need any more advice or information.

Thank you for your email. We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. Ab5439 reacts with Amphibians, Chicken, Drosophilia melanogaster, Fish, Human, S. cerevisiae, Mouse and Rat. Therefore, we will guarantee that the antibody will work with Xenopus samples in the following applications that ab5439 has been tested in: Western blot, Immunoprecipitation, Immunohistochemistry (Formalin-fixed paraffin-embedded sections), and Immunocytochemistry. Please note that ab5439 detects several members of the heat shock protein 70 kDa (Hsp 70) gene family including Hsp 70, Hsc 70, p75, and following heat shock, Hsp 72 from yeast, Drosophila, fish, mouse, avian, amphibian and human samples. As far as we are aware, cross reactivity with Xenopus has not yet been tested for use with ab5444 (so far it has only been tested for corss reactivity with human). Should you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview and in return we will award you 50 Abcam Points, which can be redeemed on a number of rewards (a further 100 points will be offered for an image). If you have any additional questions, please contact us again.