Anti-SCD1 antibody (ab23331)

I would like to test ab23331 for use on rat samples.

ANSWER:

DISCOUNT CODE: ***
Expiration date: September 11th, 2012

I am very pleased to hear you would like to accept our offer and test ab23331 in rat. This code will give you: 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for rat and 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: http://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.

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.

Please confirm that the immunogen used was indeed a synthetic peptide derived from residues 184-238 of Human SCD.

Also, has this antibody been tested against the SCD2 or SCD5 isoform?

ANSWER:

The peptide used was derived from that region - residues 184-238 of Human SCD.

The antibody has not been tested against the SCD2 or SCD5 isoform.

Please contact us again if you have any additional questions.

DESCRIPTION OF THE PROBLEM No staining

SAMPLE human liver

PRIMARY ANTIBODY ab23331 1:1000, 1:500: 1:250 overnight

DETECTION METHOD ABC

POSITIVE AND NEGATIVE CONTROLS USED human liver

ANTIBODY STORAGE CONDITIONS new fresh antibody

FIXATION OF SAMPLE Paraformaldehyde

ANTIGEN RETRIEVAL Heat mediated technique

SECONDARY ANTIBODY anti rabbit

HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

WHAT STEPS HAVE YOU ALTERED? the duration of heat mediated technique

ADDITIONAL NOTES I would like to know the standard protocol for this antibody. Do I have to perform a special antigen retrival? Is it better the enzimatic or heat mediated. At wich power and how long?

ANSWER:

I was informed that the following method was used with ab23331 immunostaining of human liver sections:

1. Deparaffinize sections in 3 changes of xylene, 5 minutes each 2. Hydrate in 2 changes of 100% ethanol for 3 minutes each, 95% and 80% ethanol for 1 minute each. 3. Rinse the sections in distilled water followed by 0.1 M PBS. 4. Quench endogenous peroxidase for 10 min with 3% H2O2 in 0.1 M PBS and wash 3 times with distilled water, 2 minutes each. 5. Retrieve epitopes by incubating the sections at 98-100° C in a microwave in 0.01 M citrate buffer (pH 6.0) for 10 minutes. Cool down in room temperature for 30-60 min. 6. Rinse the sections in 0.1 M PBS for 3 × 2 min. 7. Block the sections with 10% normal goat serum in 0.1 M PBS at room temperature for 30 min. 8. Incubate the sections with primary antibody at appropriate dilution in 0.1 M PBS overnight at 4°C. 9. Rinse the sections in 0.1 M PBS for 3 × 5 min. 10. Incubate the sections with biotinylated goat anti-rabbit IgG 11. Rinse the sections in 0.1 M PBS for 3 × 5 min. 12. Incubate the sections with HRP-conjugated streptavidin SPN-9001, Vector, 1/200 in 0.1 M PBS, 37 ° C, 20 min) 13. Rinse the sections with 0.1 M PBS for 3 × 5 min followed by 50 mM Tris-HCl buffer (pH 7.6). 14. Incubate with a peroxidase substrate solution. 15. Rinse the sections in distilled water. 16. Counterstain for 10-30 seconds with a hematoxylin counterstaining solution. 17. Rinse the sections with tap water. 18. Dehydrate through 70-80-90-95-100-100% of ethanols, each 3 minutes. 19. Clear in xylene for 2×3 min. 20. Coverslip with mounting medium. If you still experience problems with this protocol please do not hesitate to contact me again and I can offer you a refund or replacement vial if the antibody was purchased in the last 90days,