Anti-SCD1 antibody [CD.E10] (ab19862)

mouse liver samples

WB: does not see correct band at 37 kDa, but only at 26-27 kDa
Ab: 1/1000
block: milk
secondary: OK

IHC-P: nuclear localization, should be microsomal staining instead
Ab: 1/100
block: milk + goat serum
AR: citrate buffer
secondary: OK

ANSWER:

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab23331.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is notcovered by our Abpromise guarantee. Therefore, I am issuing you a testing discount code for testing this replacement antibody in mouse samples. Please see below for details.

I wish you the best of luck with your research.

===== TESTING DISCOUNT CODE =====

DISCOUNT CODE: xxx
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab23331 in mouse. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for mouseand include this 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. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: http://www.abcam.com/collaborationdiscount.

Would ab19862 react with g.pig liver tissue? If so could I have more information on your testing discount program.

ANSWER:

Thank you for calling Abcam. I have continued the search for the guinea pig sequence for stearyl-CoA desaturase, but I have not been able to locate it. I can find, rat, mouse, sheep, goat, bovine, hamster but not guinea pig. Therefore I cannot do an alignment between the human and g.pig sequences to determine likelihood of cross reactivity. If you are still interested in testing this antibody in guinea pig IHC, then I have put the testing discount details below. If there is anything else I can help with, please let me know. Testing Discount Details: As, ab19832 has not been tested in guinea pig I can offer a discount off a future purchase if you buy ab19832 now, test it in (UNTESTED APP/SPECIES) 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 discount would be to the value of 1 free (PRIMARY ANTIBODY/PROTEIN). 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 test ab19832 in guinea pig. I will then send a discount code. This code must be issued before purchasing ab19832 so please wait for my reply before ordering. 2. Purchase ab19832 either by phone, fax, or online (www.abcam.com). 3. Test it in guinea pig. 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 primary antibody 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 if ab19832 turns out to be unsuitable for guinea pig 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.  

Dear Sir/Madam,

Thanks for your email. I have tried another WB according to your suggestions about the SCD antibody. Unfortunately, the SCD antibody did not work in our lab and I agree that another

tube of antibody will not make any difference.

 I ran one WB to test the replaced SCD antibody upon I received it, I did use the same lysates I used before and I saw the similar results as before. I ran another WB with your

suggestion of using new lysates. However, I had to use the same lysate of NIH 3T3 cells, the positive control. 

We don’t have this cell line in our lab and I got one cell pellet from another lab and so now I only have cell lysate. Nevertheless, I got the similar results as all other WB generated from the first SCD antibody or the replaced SCD antibody. Please

take a look at the attached file for the results of WB. File SCD-2 is the result from the first SCD antibody with samples loaded onto lane 1-9 as NIH3T3, HepG2, Fibroblast, mouse embryonic fibroblast, mouse liver, rat liver, MCF7, neuroblastma, and Cos7. 

File SCD-4 is the result from the replaced antibody with samples loaded onto lane 1-5 as NIH3T3, HepG2, mouse liver, rat liver, and fibroblast.

 We normally make fresh lysates, then aliquot, snap-freeze, and kept them at -80oC for future analysis.

 For WB analysis, frozen lysates work fine for us and we have been doing this for a long time. Although I can’t think of a reason that a fresh lysate will make the difference, I did re-run anther WB with SCD antibody.

Unluckily, the result did not improve. However, for antibody ApoA4 I purchased from Abcam, it is another story. I didn’t saw any specific band for ApoA4 with antibody #ab108200 first. After I stripped the membrane and re-probed it with the replaced antibody

of ApoA4 (#ab81616), it gave me the band at the right size and the background was clear.

 After so many WB and such a long time trying, I have to say that it clearly does not work in our lab. I was wondering if it is possible that we could get a refund for this antibody.

I would greatly appreciate if you could consider the possibility.

 Best Regards,

ANSWER:

Thank you for your email. We highly appreciate your cooperation in this.

I am sorry we are unable to resolve the problems you have been experiencing. It may be possible that these antibodies are not suitable for Typhoon method with Kpi lysis buffer. The ab19862 was cited in 6 publications as far as we know and in 2 publications PubmedID 19787047 and 18697866 in WB so I might suggest checking the protocol they have used.

I have asked our account department to refund the money on the original order 912415. The refund ID is 17800. Please contact our customer service team by quoting this order in case of any question. You can also inform your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

Dear Sir/Madam,

I appreciate your help on this matter and I have received the new ab19862 last Friday. I checked the tube and it is from a different batch. I have

run a WB with 9 different cells and tissues on one membrane as I have used to test the previous antibody. I have run it at the same condition and the dilution for primary antibody is 1:500 (v/v). Unfortunately, I am still unable to get SCD band on all these

samples. I got a similar result just as before with the previous antibody.

I really have hoped that this would be the last email to you and I could have started my project after this.

Unfortunately, it is not the case. Based on the website of Abcam, ab19862 would be the best option for our project since we are going to run some WB with cell lines and tissues from human, mouse, and rat. I was wondering if anybody else also ordered

this antibody recently and if it worked for them. I just want this WB to work.

Thanks for help me on this matter and I am sorry to bring you the bad news.

Best Regards,

ANSWER:

Thank you for contacting us.

The free of charge replacement is also covered by Abpromise guarantee so we can certainly help with your requirements. I am very sorry to hear; the result did not improve.

I am however not sure whether the problem might be due to antibody, lysates or experiment itself. As per your email you have used the protocol successfully with other antibodies I believe the protocol is fine. However I would like to ask whether you have prepared different lysates or used same lysates as you used before. If you have used same previous lysates then I would suggest preparing fresh lysates and using the antibody again.

We have sold many units of this antibody and we have not received many complaints that can ring the bell regarding antibody specificity. I also feel that sending another free will not be making any difference so I would suggest trying different lysis buffer and fresh lysates.

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

Sample: Mouse Tissue sections (Skin sections) Application: Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) Blocking step:: BSA as blocking agent for 30 mins  Concentration: 3% Fixation:: Paraformaldehyde Antigen retrieval step:: Heat mediated Incubation time:: 12 hours  Positive control:: MOUSE ANTIBODY TO P63 Negative control:: MOUSE ANTIBODY TO P63 - I.E. STAINS SOMETHING DIFFERENT Dilution: 1/100 Rating: No detection - I would not use again Additional notes: SECONDARY: 1:100 ANTI-MOUSE FAB FRAGMENT ALEXA 568 DETECTED SOME OTHER CELL TYPE (?MAST CELL) BUT NOT SEBACEOUS GLAND WITH THIS ANTIBODY. MOUSE ANTI-P63 (ABCAM) WORKED USING THE SAME SECONDARY. Secondary antibody: Name:: Non-Abcam one was used: ANTI-MOUSE FAB A11019 Conjugation: Alexa Fluor® 568 Dilution:: 1/100 Abcam response: The product ab19862 has not been tested in IHC-P and the results presented here suggest that this antibody should not be used in this application. However, it is possible that optimization of this protocol could give a more positive result.

ANSWER:

Thank you for submitting an Abreview - ReviewID: 6368 and AbID: 19862 .

We very much appreciate your honest feedback about this product and have published it to help future users of this antibody as our policy is to publish negative and positive feedback and reward both types with Abcam Points.

However, I would like to point out a couple of things:

1) IHC-P is an untested application and ab19862 might not work in IHC-P or your protocol may need to be optimized.

2) Have you tried troubleshooting e.g. testing different antigen retrieval methods

Below please find a link to our troubleshooting questionnaire. This will give us further information regarding the samples you tested and the protocol you used. Once you have submitted the form to us, we will look at the protocol you used and see if there are any suggestions we can make to improve your results.

The questionnaire can be found at:

http://www.abcam.com/index.html?section=ihc&pageconfig=technical&intAbID=19862&mode=questionaire

I look forward to receiving your reply.