HeLa全细胞裂解物(ab29545)

Q&A (13)

概述

  • 产品名称

    HeLa全细胞裂解物
    参阅全部 HeLa 细胞裂解液

  • 常规说明


    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.

  • 经测试应用

    适用于: WBmore details

性能

  • Mycoplasma free

    Yes

  • 形式

    Liquid

  • 存放说明

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

  • 存储溶液

    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...

  • 裂解物说明

    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.

  • 研究领域

  • 背景

    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.

应用

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.

应用 Ab评论 说明
WB Use at an assay dependent dilution. Samples need to boiled for 5 minutes and put on ice before use.

实验方案

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

文献 (0)

发表研究结果有使用 ab29545？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab29545 尚未被引用在任何文献中。

客户评价及客户问答

Q&A
提交问题

1-10 of 13 Q&A

Question

We are currently using LAT1 ab85226 for Western blotting, but the signal is really low (even when we have increased the amount of Ab). I was thinking of switching to ab134121 instead. Would we still be measuring the same LAT1 protein? We are measuring it in human muscle. I have a couple questions (enumerated for easy reference):

1) Is this an appropriate application of the RabMAb?

2) What secondary Ab do we need to use?

3) We standardize everything to a rat standard. The RabMAb do not react with rat, correct? I need to buy ab89545 as a positive control? Can I still standardize my unknowns to HeLa positive controls?

4) Realistically, what dilution would we be using? (I want to make sure I order enough from the same lot number)

I apologize if some of these questions are basic; Western blotting is not my strong point.

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Answer

Thank you for contacting us.

Yes, both ab85226 and ab134121 detect the same SLC7A5 (LAT1) protein. http://www.uniprot.org/uniprot/Q01650

We would guarantee ab134121 to work in WB, IP, and IHC-P with human samples. We know that it does not work well with Flow Cyt, ICC/IF and rat or mouse samples.

Cat # ab89545 is an EEA1 antibody no longer available on our catalogue. You must mean ab29545, the HeLa cell lysate. Yes, that would be a good positive control for use with ab134121.

In WB with ab134141, we recommend a working dilution of 1/1000 - 1/10000. The amount that you will need will depend on the amount present in your samples. The image generated on our website was done with a 1/1000 dilution for example.

A suitable secondary would be Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) to be used with ECL detection, like our ab133406.

https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rabbit-IgG-HL-HRP-ab97051.html

https://www.abcam.com/optiblot-ecl-detect-kit-23pg-187ng-ab133406.html

You can find our full range of Optiblot WB reagents here: https://www.abcam.com/index.html?pageconfig=resource&rid=14709&source=pagetrap&viapagetrap=optiblot

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

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Question

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 to use it in my experiments.  So if you could send those to me that would be great. 

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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 am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxxxx. Included in this order are positive control lysates for these products. 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.

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Question

Dear recipient,
Does the Dkk1 antibody ab 61275 include the positive control mentioned
in the data sheet (HeLa cell lysate)? Or should I purchase it
separately?
Thank you very much for your reply.
Best regards,

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Answer

Thank you for contacting us.

I can confirm that the positive control is not included into an order of the DKK antibody ab61275.

However, we do offer two products of HeLa cell lysate: ab29545 (tested for WB application) and ab150035 that can be used as positive controls for WB. For further information about both products use the provided links below.

ab29545
https://www.abcam.com/index.html?datasheet=29545 (or use the following: https://www.abcam.com/index.html?datasheet=29545).


ab150035
https://www.abcam.com/index.html?datasheet=150035 (or use the following: https://www.abcam.com/index.html?datasheet=150035).


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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question

We have just received HeLa Whole Cell Lysate (ab29545). According to datasheet it is recommended to store this product at -80°C but unfortunately the product arrived at room temperature . Only after one day ab29545 was placed to -80°C.

We would like to ask if this kind of conditions does not impaired the activity of the product?

Thank You in advance for Your answer

Warm regards

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Answer

Thank you for contacting us.

We ship many products at 4C so it is not uncommon for this product to be arrived at room temperature. The product will be absolutely fine, please use it without any hesitation.

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question

phone call: - parcel has been left at RT

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Answer

Merci de nous avoir contactés.

Comme discuté au téléphone, nous allons vous envoyer un remplacement gratuit du lysat cellulaire HeLa ab29545. Jeme permets de vous transférer le numéro de commande de remplacement gratuit de ce produit :xx . Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition.

N'hésitez pas à nous contacter lors d'une prochaine occasion.

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Question

I have a question related to that previous order.

The lysates I bought are not ideal for my purpose, the PEG content is too high for LCMS analysis. And I am afraid of how much protein loss I will incur if trying to remove them from the sample. Would it be possible to get the same cells (regular, IL1 and TNF stimulated) but in a pre-lysate form (PBS washed pellets or lyophilized form) so that I could carry my own sample prep?

I would be interest in i.e. 10-100 million cells quantity.

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Answer

Thank you for contacting us.

We do not sell cells, only cell lysates. I apologize but we will not be able to provide these in a pre-lysate form.

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question

Product code: 2954
Inquiry: Dear All I am looking for a Cell Line . Is Ab29545 is the right products...? Thank you

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Answer

Thank you for contacting us.

We unfortunately do not sell cell lines.

The product ab29545 is a cell line lysates for use in western blot.

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question

WB with mouse small intestine mucosa
sees bands at 55 and 70 kDa
Ab from other company also shows bands at 55 and 70, but also 120 kDa
Ab: 1/200 o/n
block: 5% milk
2nd: HRP, works well
suggested Hela lysate as positive control as blocking peptide is not available

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Answer

Thank you for contacting us.

I have arranged for the shipment of the Hela cell lysate ab29545 on order number xxx.
Please let me know what results you are obtaining and if this helps to interprete your results with the mouse samples.

I look forward to hear back from you and assist you further.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question

Either no signal in WB, or seeing band of interest, plus a non-specific band.

Read More
Answer

Thank you for calling Abcam.

I have sent you a vial of ab29545, HeLa cell lysate to use as a positive control.

Please let me know if there is anything else I can help you with, or if continue to experience any issue with our antibodies.

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Question

I have used ab29545 for western blotting with a loading control anti tubulin antibody and there was no signal. I used 3 other cell lysates I purchase from Abcam with the same antibody and they all worked as expected.

Read More
Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab29545.

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.

Read More

1-10 of 13 Q&A

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