Anti-Angiotensin Converting Enzyme 1 antibody [2E2] (ab11734)

Hello,
What is it better to use as a positive control in Western Blot: a
tissue lysate or a recombinant purified protein ?
Thanks for your answer

ANSWER:

Thank you for your response and the new enquiry.
This may depend on the actual antibody itself. If the antibody is raised against a full length or a fragment of recombinant protein, then it would be advisable to use rec. protein as positive control.
For this particular antibody, denatured human kidney angiotensin converting enzyme was used as immunogen (as the datasheet indicates) so tissue lysates containing the target can be applied as good and recommended positive control.
This clone recognises denatured ACE, binding to an epitope in the C domain, and it has been reported to be active in Western blotting.
I hope this helps and if I can assist further, please do not hesitate to contact me.

LOT NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM I am looking for a positive control to use with this antibody. I am doing western-blot with sheep protein lysate. SAMPLE sheep protein lysate PRIMARY ANTIBODY ab11734 SAMPLE PREPARATION lysis buffer from Q proteome kit, laemli buffer + boiled 10 min TRANSFER AND BLOCKING CONDITIONS 5% non fat dry milk SECONDARY ANTIBODY ab97265

ANSWER:

Thank you for your enquiry regarding ab11734.
Angiotensin Converting Enzyme 1 (ACE 1) is expressed with highest levels in lung, kidney, heart, gastrointestinal system and prostate. Tissue lysates from any of these tissues would be a good positive control for detecting ACE 1. I would advise you to use RIPA buffer containing a mixture of different proteinase inhibitors to prevent any protein degradation. The recipe of this buffer can be found at our website:
http://www.abcam.com/index.html?pageconfig=resource&rid=11379#A1
If you need any further assistance in the future, please do not hesitate to contact me.

I am using ACE Anti-Angiotensin Converting Enzyme 1 antibody [2E2] (ab11734), and Anti-WIF1 (ab105925) both of these are not quantified. Can you let me know the total protein concentration in each of these please.

Best Wishes,

ANSWER:

Thank you for your recent enquires and for your patience.

I now have the following information from the originators of these antibodies regarding the total protein concentration. I am sorry we do not have an accurate concentration, but they have been able to provide an estimate. As discussed on the telephone, this is total protein concentration, and not the antibody concentration:

ab11734 Angiotensin Converting Enzyme 1 antibody [2E2] 

' Unfortunately, neither we nor the originators routinely assay for the concentration of this product. Tissue culture supernatant formats would normally contain about 10 to 50ug/ml of antibody but we are unable to be more specific than this.'

ab105925 WIF1 antibody [1G5] 'The total protein concentration is approximately 5 mg/ml.' I hope this will be helpful to you.

If you have any further questions, please do not hesitate to contact us.

Please can you let me know the concentrations of Anti-WIF1 (ab105925) and Anti-Angiotensin Converting Enzyme 1 antibody [2E2] (ab11734).

ANSWER:

THank you for contacting us. Ab105925 is from ascites and ab11734 is from tissue culture supernatant. Concentrations are not determined for these types of products as they are not relevant unless the antibody is purified, as they may contain proteins other the target. We refer you to the recommended dilutions for your application which can be found on the datasheet. Please let me know if you have any further questions.

Can you please let me know if the epitope is C or N term? Is it also possible to find out the size for this product?

ANSWER:

Thank you for your enquiry. This clone recognizes denatured ACE, binding to an epitope in the C domain. The predicted molecular weight: 170 (somatic) or 90 (germinal) kDa.

Please contact us again if you have any additional questions.