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HeLa whole cell lysate (ab29545)

Submit a review Q&A (13)References (2)

Overview

  • Product name

    HeLa whole cell lysate
    See all HeLa lysates

  • General notes


    Protein is isolated from HeLa cell pellet using a proprietary technique. The pellet was frozen in liquid nitrogen immediately and then stored at -70ºC. Quality control: 1. The isolated protein pattern on SDS-PAGE gel is visualized by coomassie blue staining. The pattern is consistent with each lot. 2. The isolated protein is Western Blot analyzed by either a GAPDH or beta-actin antibody. The expression level is consistent with each lot.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Mycoplasma free

    Yes

  • Form

    Liquid

  • Storage instructions

    Allow to warm to room temp and agitate gently before aliquotting. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.

  • Storage buffer

    Constituents: 1% 4-Nonylphenol, branched, ethoxylated, Potassium chloride, 0.1% Bromophenol blue, Magnesium chloride, 1.2% Beta mercaptoethanol, 2% Sodium lauryl sulfate, HEPES, Sucrose, 10% Glycerol

    With Protease inhibitor cocktail/EDTA/Sodium deoxycholate
  • Concentration information loading...

  • Lysate notes

    Protein is isolated from HeLa cell pellet using a proprietary technique. The pellet was frozen in liquid nitrogen immediately and then stored at -70ºC. Quality control: 1. The isolated protein pattern on SDS-PAGE gel is visualized by coomassie blue staining. The pattern is consistent with each lot. 2. The isolated protein is Western Blot analyzed by either a GAPDH or beta-actin antibody. The expression level is consistent with each lot.

  • Research areas

  • Background

    HeLa 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

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab29545 in the following tested applications.

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

Application Abreviews Notes
WB
Use at an assay dependent dilution. Samples need to boiled for 5 minutes and put on ice before use.
Notes
WB
Use at an assay dependent dilution. Samples need to boiled for 5 minutes and put on ice before use.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (2)

Publishing research using ab29545? Please let us know so that we can cite the reference in this datasheet.

ab29545 has been referenced in 2 publications.

  • Vonk PJ & Ohm RA H3K4me2 ChIP-Seq reveals the epigenetic landscape during mushroom formation and novel developmental regulators of Schizophyllum commune. Sci Rep 11:8178 (2021). PubMed: 33854169
  • Meng H  et al. L450W and Q455K Col8a2 knock-in mouse models of Fuchs endothelial corneal dystrophy show distinct phenotypes and evidence for altered autophagy. Invest Ophthalmol Vis Sci 54:1887-97 (2013). PubMed: 23422828

Customer reviews and Q&As

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11-13 of 13 Abreviews or Q&A

Question

Thank you for your prompt answer. 1) How have you prepared the samples: which lysis buffer have you used, have you denatured them by boilingand how have you applied the Abcam controllysates (HeLa, prostate whole tissue)? (It would be great if you could provide the catalog numbers, lot numbers and/or order numbers/dates for these). Samples preparation (my samples and those from ABCAM): I prepared the loading buffet with 950ul of Laemmli Sample Buffer (BIORAD) and 50ul of Betamercaptoethanol. I mixed 25ug of sample + water (to reach 15ul) + 14ul of loading buffer + 1ul of DTT. I boiled them for 5 minutes at 95°C then loaded them. 2) Could you please confirm the blocking agent used (milk or BSA)? I used Superblock T20 blocking buffer 3) At which dilution and which incubation temperature/time have you applied the primary antibody? dilution of 1:1000, incubation at 4°C overnight 4) Have you run a no-primary control to check the secondary antibody? No because we routinely use this secondary antibody and we never had a problem. I and other colleagues have used the same lot I used in this experiment the following day with another primary antibody and it worked very well. It is part of the ECL Western Blotting Analysis Kit. Once we have received this information and it turns out that the antibodyis not functioning as cited on the product data sheet, we will be pleased to provide afree of charge replacement, acredit note or refund.

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Answer

Thank you for confirming these details. The details you have kindly provided will provide us with vital information for our monitoring of product quality. Although it is not necessary to add further loading buffer to the lysates you purchased from us, I do not see a reason why it should not work the way you processed the samples. Also, I am pleased to hear that the secondary antibody works fine - that was all we needed to know about it. Thus, reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably received a bad vial. I apologise for the inconvenience and am pleased to offer you a free of charge replacement with a new vial of ab87811. Otherwise, as we currently do not have an alternative Anti-DAB2IP antibody available, I could send you any primary antibody of your choice or issue you a credit note in compensation. Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Question

Are there nucleic acids present in this lysate, or is it mostly just protein?

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Answer

Thank you for your enquiry. This lysate doesn't contain nucleic acids. We tested this by PCR before. If there are nucleic acids, it is a very small amount of DNA which could only be detected by more sensitive methods. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Question

Would HeLa cell lysate ab29545 be ok as WB positive control for Ki67 antibody ab16667? Need to find out if HeLa cells express Ki67. If not, what else can be used as a positive control? Tel 0208 8693375

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Answer

Thank you for your telephone enquiry. I am sorry for the delay in responding. HeLa cells do express Ki67 (shown on the data sheet for Ki-67 antibody ab1550 tested on HeLa cells). HeLa whole cell lysate (ab29545) will therefore be a suitable Ki67 positive control for your experiments. We also have a Ki67 peptide available, ab15581. I hope this is helpful. Good luck with your research! Please do not hesitate to get back in touch if you have any more questions. I will be happy to help.

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11-13 of 13 Abreviews or Q&A

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