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HeLa (Human epithelial carcinoma cell line) Nuclear Lysate (ab14655)

Q&A (10)Specific References (1)

Overview

  • Product nameHeLa (Human epithelial carcinoma cell line) Nuclear LysateSee all HeLa lysates ...
  • General notesWe recommend aliquoting the extracts into single-use fractions and then storing them.
  • Tested applicationsWB, EMSA more details

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferpH: 7.90
    Constituents: 0.48% HEPES, 20% Glycerol, 0.01% Magnesium chloride, 0.077% DTT, 0.004% EGTA, 2.05% Sodium chloride, 1% NP-40
    Note: With Protease inhibitors
  • Concentration information loading...
  • Research Areas
  • BackgroundHeLa cells are human epithelial cells from a fatal cervical carcinoma. The cell line was derived from cervical cancer cells taken from Henrietta Lacks, in 1951. Horizontal gene transfer from human papillomavirus 18 (HPV18) to human cervical cells created the HeLa genome which is different from either parent genome in various ways including its number of chromosomes. HeLa cells have a modal chromosome number of 82, with 4 copies of chromosome 12 and 3 copies of chromosomes 6, 8, and 17. HeLa cells are adherent cells (they stick to surfaces) and maintain contact inhibition in vitro.

    • Applications

    Our Abpromise guarantee covers the use of ab14655 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Notes
    WB WB: Use at an assay dependent dilution.
    EMSA EMSA: Use at an assay dependent dilution. PubMed: 12075007

    References for HeLa (Human epithelial carcinoma cell line) Nuclear Lysate (ab14655)

    This product has been referenced in:
    • Weitzel LR  et al. Discovery and verification of protein differences between Er positive/Her2/neu negative breast tumor tissue and matched adjacent normal breast tissue. Breast Cancer Res Treat 124:297-305 (2010). WB . Read more (PubMed: 20087651) »

    See 1 Publication for this product

    Publishing research using ab14655 ? Please let us know so that we can cite the reference in this datasheet

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    Submit a Question for ab14655

    Displaying 1 - 10 of 10 results for Q&A

    10
    Votes: Highest First

    I am more than happy to try a new vial of p53.  Do you also have a positive control for this antibody that I can also use?  Also the LSS positive control would be very helpful because if that band is truly LSS then that is a good antibody and I would like...

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    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...

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    Hello, I’m sending pictures of my blots because I’m unsure of what band is my protein of interest. Can you point out the correct band in my blots? The following is a description of what I did: Samples The samples are lysate fractions...

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    Thank you for contacting us.

    I would like to thank you for sending me your protocol, this information is a great help.

    I did not recieve any images with you email however. Could you please resend those to me?

    I hope this...

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    Hello,



    I’m sending pictures of my blots because I’m unsure of what band is my protein of interest. Can you point out the correct band in my blots?



    The following is a description of what I did:

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    Thank you for contacting Abcam.

    I have attached the ppt slide with a red arrow next to the bands that I think are the correct bands(they are the same that you have marked).

    There are a few protocol changes that I would recommend to red...

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    Hello Technical Support,

    Perhaps a refund would be best as I am interested in the nuclear lysate.

    Thank You,

    Thank you for your reply.

    I am sorry that we were not able to resolve the issues you were having with ab14655 and I have processed your request for a refund. Your credit note number is ******.

    Please let me know if there is anything el...

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    Thank you for your prompt reply. Please send the HeLa nuclear lysate that you described; just to confirm it is a nuclear lysate and not a cell lysate? I appreciate your help and patience with this matter.

    Thank you for your reply.

    The product ab29545 is a whole cell lysate not a nuclear lysate. The only nuclear lysate we have in stock is the ab14655that you currently have.

    If you still wish for me to send you ab29545, the whole cell lys...

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    I must have misunderstood you, I loaded 18uL per well (I had 10uL of HeLa nuclear lysate and added 10uL of 2x protein loading buffer to a total volume of 20ul of which I loaded 18ul per well). The wells themselves if loaded carefully can hold 22uL of samp...

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    Thank you for that information.

    I do not think adding 4ul more will increase your signal and since you can only load a maximum of 22ul/well, I would like to send you ab29545 as a free of charge replacement. This HeLa cell lysate is already in sa...

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    I loaded 2.5mg/ml of total protein (the original concentration) I also tried lower concentrations (0.5 mg/mL and 0.25 mg/mL) and saw signal only in the PC12 nuclear lysate of corresponding concentrations. I assume that if the PC12 lysate showed a clean si...

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    Thank you for your reply.

    I appreciate your patience in this matter, but when you say you loaded 2.5mg/ml of total protein, how many microlitres of lysate is that? I ask, as we send 100ug of total protein at at concentration of 2.5mg/ml (therefo...

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    On January 10, 2012 I placed an order for HeLa and PC12 nuclear lysates (order confirmation # **********). I ran both lysates at 2.5 mg/ml (original concentration) and stained the blot with three different antibodies; among them with a anti-TATA binding p...

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    Thank you for contacting Abcam.

    I am sorry about the problems you have been having with ab14655, HeLa cell lysate. I was wondering if you could tell me how much total protein you loaded of the cell lysate. I ask as there does seem to be a signal...

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    Thanks for your letter, however you missunderstand me, most probably it is my mistake. I'd like to know how you, at Abcam, are preparing nuclear extract e.g. to produce HeLa nuclear ectract cat# 14655 or Hep G2 nuclear extract cat# 14660. How do you get...

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    Thank you for your enquiry. The nuclear extracts were prepared as detailed in: Dignam, J.D., Lebovitz, R.M. and Roeder, R.G. (1983) Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucl. Aci...

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    Whould you please inform me about the procedure you use to prepare nuclear extracts (e.g. form HeLa cells). Thanks!

    Thank you for your enquiry. To prepare a nuclear lysate from HeLa cells I would recommend that you use RIPA buffer as follows. RIPA buffer (RadioImmunoPrecipitation Assay) buffer: RIPA buffer contains the ionic detergent sodium deoxycholate as an ...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"