A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)

If you do not have ovary cell lysate at present, the human testis tissue lysate will do, too.(http://www.abcam.com/Testis-Human-Tissue-Lysate-adult-normal-tissue-ab30257.html). Thank  you very much!

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 have issued a free of charge control lysate (ab30257) as goodwill with the order number.

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 also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

I have read the two papers you mentioned about, but I still do not think they are very convincing. First, the  GnRH receptor is different from  HCG receptor either in  function or in structure. Even the existence of  GnRH receptor in HepG2 cells does not mean there are HCG receptors in them, too. And, although there are some changes when HepG2 cells being cultured with hCG, there is no direct proof that the HCG receptor is definitely there. While in the paper I mentioned(    Shan LX, Hardy MP  Endocrinology. 1992 Sep;131(3):1107-14.  ( PMID:  1505454,  ),  the authors did the Northern blot analysis showing there was no HCGR mRNA in HepG2 cells. I think this is a more solid proof. I want to know whether your company has ever tested the Anti-HCGR antibody on HepG2 cells . Because in my experiment, the Western blot showed some bands in HepG2 lane, but not at  79KDa band weight which is showed in your protocol. I do not know whether it is caused by the lack of specifity of this antibody. If you have tested the antibody in HepG2 cells, I want to know whether the results are the same. If you have not tested it, I do not think it is a good idea to list the HepG2 cells in the positive control before more convincing results been published. Thank you very much! Best wishes,

ANSWER:

Thank you for taking the time to look into this and for your response.

We have not tested this in HepG2 cells and the positive control recommendations were based on the literature. I thank you for your recommendation with this and have removed HepG2 from the controls.

Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Phoned because of concern regarding the positive controls listed on the datasheet. According to published reference, HepG2 is a negative control.

ANSWER:

Thank you for contacting us.

I was able to find the reference that you spoke of on the phone (ShanLX, HardyMP Endocrinology.1992 Sep;131(3):1107-14. (PMID:1505454,http://www.ncbi.nlm.nih.gov/pubmed/1505454).

During my research for this question I was able to find a later paper in whichHepG2 lines are shown to haveGnRHreceptors,Pati D,Habibi HR.Endocrinology.1995 Jan;136(1):75-84.(PMID:7828560; ).

A more recent paper, Jacobson HI, et.al.Adv Exp Med Biol.2008;617:477-84. (PMID: 18497072,http://www.ncbi.nlm.nih.gov/pubmed/18497072) use HepG2 culture with hCG as well. Is seems that the early information has been supplanted by this new evidence. Therefore I believe that the HepG2 cells should be a positive control for this product.

If you have further concern I would recommend using A431 cell lysate (http://www.abcam.com/A431-Human-epithelial-carcinoma-cell-line-Whole-Cell-Lysate-ab7909.html) or H1299 cells.

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

What would be a suitable positive control for this product 14-3-3 (phospho S) antibody (0014-33)?

ANSWER:

Thank you for your enquiry.

For immunohistochemistry paraffin-embedded sections, human breast carcinoma was used in as control. For Western Blot QC, lysates were prepared from A431 cells, untreated or calyculin A-treated. A431 cells are a human epithelial carcinoma cell line. Calyculin treatment greatly increased the expression of the 14-3-3 Binding Motif, phosphorylated (Ser) in a number of different proteins.

Abcam lists this product ab7909, A431 (Human epithelial carcinoma cell line) Whole Cell Lysate, which could serve as a decent positive control for Western Blotting.

Hi, I am interested to buy the lysate A431+EGF but it is not to do western blot, it is to do some immunoisolation of phosphoproteins. So I don't want the lysate in SDS-Page buffer. I want it in lysis buffer. Do you tell me if you have this kind of lysate and if not where I can find it. Thanks, Manon

ANSWER:

Thank you for your enquiry and your interest in our products. Unfortunately, we cannot change the constitution of the storage buffer for this product (Ab7909: A431 whole cell lysate). If you cannot find what you are looking for within the Abcam catalog, I suggest that you click on the link to The world's Antibody Gateway. The world's Antibody Gateway is a free search engine service provided by Abcam to help you to quickly find the antibodies that you are looking for. It is a free-text search engine developed by the Abcam team so that it searches the catalogs of all online antibody companies (currently 249).