Anti-CD10 antibody [56C6] (ab951)

Product code: 951
Lot number: GR79429-1
Inquiry: How should the Renoir Red solution be used in conjunction with the CD10 antibody? Also, no concentration was listed on the tube or the product sheet, although it was specified that the antibody should be used at 5 ug/ml. What is the concentration of the 250 ul of CD10 antibody (ab951)?

ANSWER:

Thank you for contacting us.

The IgG concentration for lot GR79429-1 is 7.2µg/mL. For IHC, the antibody should be used at a concentration of 1/20 - 1/50. The antibody was validated for IHC with a sensitive polymer detection system, so if you are using a different kind of detection, for instance an ABC kit, or a conjugated secondary without any ABC or polymer amplification, we recommend the lower dilution of 1/20. A bottle of the diluent Renoir Red is included as the optimal diluent. Just substitute it in place of your standard buffer diluent.

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

LOT NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Multiple bands. The expected molecular weight is 85 kDa, whereas I got a major slower band at about 100 kDa (which I believe is the correct band) and a faster, minor band at 75 kDa. Does the protein degrades/alterates upon boiling? Samples were, as usual, boiled for 3' before loading. SAMPLE Mouse brain extract. PRIMARY ANTIBODY AbCam ab951 DETECTION METHOD ECL and X-ray film ANTIBODY STORAGE CONDITIONS Antibody was supplied already dissolved and was stored at 4°C, as indicated. SAMPLE PREPARATION Fractionated extraction using: Fraction 1) Tris-HCl buffer, aprotinin, leupeptin, NP40, SDS, PMSF, Na3VO4, EDTA. Fraction 2) TNT buffer Fraction 3) Same as buffer 1, but with the addition of deoxycolate and EGTA in place of EDTA. AMOUNT OF PROTEIN LOADED 10 micrograms. ELECTROPHORESIS/GEL CONDITIONS Reducing 4-12% Bis-Tris gel, XT-MES BioRad buffer. TRANSFER AND BLOCKING CONDITIONS Used the TurboBlot transfer system by BioRad on nitrocellulose membrane. Blocking 4% non-fat dry milk in 0.2% Tween-TBS for 2h at R/T. SECONDARY ANTIBODY Jackson HRP-conjugated goat anti-mouse HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

ANSWER:

Thank you for contacting Abcam.

I think you are correct that the band that you are seeing at 100kDa is correct as based on the Swissprot sequence http://www.uniprot.org/uniprot/Q61391, there are q few glycosylation sites on this protein that would account for the increase in the predicted molecular weight. The 75kDa mat be a non-specific band in which case changing your blocking reagent to 5%BSA may help to reduce that signal. Otherwise if it is a breakdown product then you could increase the amount of protease inhibitors that you are using.

The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot on mouse samples . If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

Please let me know if there is anything else I can help you with.

We recently ordered anti-CD10 antibody [56c6], ab951. The antibody arrived with 25 mil of renoir red diluent. What is the purpose of the diluent? The antibody is already suspended in solution and the product information sheet offers no information about the renoir red diluent except that it is in the storage buffer for the antibody.
Thank you for your help.

ANSWER:

Thank you for contacting Abcam.

Renoir Red Diluent is specifically formulated to enhance certain types of antibodies and may increase titers up to 2 to 4 times. The diluent is a colored-coded red for identification. It is specially designed to work with Tris-based antibodies, rinsing buffers and all detection systems. It can also be used on an automated staining system. You do not have to use this product, we only ship it with this antibody as the storage buffer it is in, is Renoir Red.

Please let me know if there is anything else I can help you with.

Dear Sir/Madam Please kindly recommend the abcam slide control for IHC-P application for this antibodies. 1. Anti-ErbB 2 antibody (ab2428) 2. Anti-Progesterone Receptor antibody [SP2] (ab16661) 3. Anti-Estrogen Receptor antibody [SP1] (ab16660) 4. Anti-Myeloperoxidase antibody (ab45977) 5. Anti-TdT antibody (ab85148) 6. Anti-TMEM16A antibody (ab53212) 7. Anti-NSE antibody [EPR3377] (ab79757) 8. Anti-CD23 antibody [EPR3617(IHC)] (ab92495) 9. Anti-CD10 antibody [56C6] (ab951) 10. Anti-Melanoma antibody [HMB45 + DT101 + BC199] (ab732) 11. Anti-CD79a antibody [HM47/A9] (ab3121) 12. Anti-CD45RO antibody [UCHL-1] (ab86080) 13. Anti-CD45 antibody (ab10559) 14. Anti-CD30 antibody [CON6D/ C2] - BSA and Azide free (ab76518) 15. Anti-CD15 antibody [(Workshop IV) (BRA-4F1) our designation 238)] (ab115993) Best regards

ANSWER:

Thank you for your enquiry. I am pleased to provide the following suggested positive controls for these antibodies. I have mainly chosen tissue that has been tested succesfully and shown in an image on the datasheets: 1.Anti-ErbB 2 antibody (ab2428) Human breast cancer 2.Anti-Progesterone Receptor antibody [SP2] (ab16661) Human breast carcinoma 3.Anti-Estrogen Receptor antibody [SP1] (ab16660) Human breast carcinoma 4.Anti-Myeloperoxidase antibody (ab45977) Human spleen tissue 5.Anti-TdT antibody (ab85148) Human testis 6.Anti-TMEM16A antibody (ab53212) Human liver 7.Anti-NSE antibody [EPR3377] (ab79757) Human brain tissue 8.Anti-CD23 antibody [EPR3617(IHC)] (ab92495) Human tonsil 9.Anti-CD10 antibody [56C6] (ab951) Lymph node tissue or human breast microepithelium 10.Anti-Melanoma antibody [HMB45 + DT101 + BC199] (ab732) Murine melanoma metastasis tissue 11.Anti-CD79a antibody [HM47/A9] (ab3121) Human Lymphoid tissue, tonsil 12.Anti-CD45RO antibody [UCHL-1] (ab86080) Human Tonsil, Thymus 13.Anti-CD45 antibody (ab10559) Human Tonsil tissue 14.Anti-CD30 antibody [CON6D/ C2] - BSA and Azide free (ab76518) Human Hodgekins lymphoma tissue 15.Anti-CD15 antibody [(Workshop IV) (BRA-4F1) our designation 238)] (ab115993) Human Hodgekins lymphoma tissue Human spleen I am providing a list ofslides that we sell which I hope will be helpful to you. I can recommend to review the online datasheet for more details. Human brain: ab4305Brain (human): hippocampus normal tissue slides http://www.abcam.com/index.html?datasheet=4305 (or use the following: http://www.abcam.com/index.html?datasheet=4305). Human lymph node: ab4350Lymph node (human) normal tissue slides IHC-P, ISH http://www.abcam.com/index.html?datasheet=4350 (or use the following: http://www.abcam.com/index.html?datasheet=4350). Human testis: ab4373Testis (human) normal tissue slides IHC-P, ISH http://www.abcam.com/index.html?datasheet=4373 (or use the following: http://www.abcam.com/index.html?datasheet=4373). Human liver: ab4348Liver (human) normal tissue slides IHC-P, ISH http://www.abcam.com/index.html?datasheet=4348 (or use the following: http://www.abcam.com/index.html?datasheet=4348). Human spleen: ab4372Spleen (human) normal tissue slides IHC-P, ISH http://www.abcam.com/index.html?datasheet=4372 (or use the following: http://www.abcam.com/index.html?datasheet=4372). I am sorry we do not have slides forthe following: Human tonsil Hodgekins lymphoma tissue Mouse melenoma tissue I hope this will be helpful to you. If you have any further questions,please do not hesitate to contact us.

Hello,

We have recently received a consignment of antibodies from you on Friday. Unfortunately, I was not informed of this delivery until today and therefore the antibodies have been left at room temperature over the weekend. I have now placed the antibodies immediately into 4C for storage but am wondering if these antibodies would still be viable after being left at room temperature all this time. These antibodies are unconjugated antibodies (both monoclonal and polyclonal). I would appreciate if you can let me know as soon as possible. Many thanks.

Regards,

ANSWER:

Thank you for contacting us.

Abcam takes product quality seriously and we recently undertook an in-depth study examining storage conditions and antibody performance. After over 13,000 individual ELISA experiments, we have determined that storing our antibodies at various temperatures of up to 45°C for 1 week does not impact their activity. I would be happy to share a summary of our findings with you; please let me know if you would like to see this.

As a precautionary measure only, we ship our antibodies in packaging with ice packs to provide extra temperature stability in transit. The ice pack may be thawed or even at room temperature when you receive it; please be assured that this is normal and that your product is safe to use. Once you have received the vial, please follow the long-term storage instructions on the datasheet.

Our Abpromise to you is that, should our product not work as stated on the datasheet, we will resolve the issue to your satisfaction, either with helpful advice to optimize your experiment or a replacement or refund of the product. Please contact our knowledgeable Scientific Support staff should you experience any issues while using our products. Contact information can be found at:

http://www.abcam.com/index.html?pageconfig=contactus

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

PS: Attached is the tested data of few of our antibodies.