BALB/3T3 whole cell lysate (ab7901)

Q&A (2)References (1)

Overview

  • Product name
    BALB/3T3 whole cell lysate
  • General notes
    Cell line: BALB/3T3 clone A31 (Mouse embryonic fibroblast).
    Growth media: DMEM and 10% FBS (Fetal bovine serum).


    BALB/3T3 clone A31 cell lysate was prepared by homogenization in modified RIPA buffer(150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl flouride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 µg/ml of aprotinin, 5 µg/ml of leupeptin). Cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The cell lysate was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol.

  • Tested applications
    Suitable for: WBmore details

Properties

  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Constituent: 5% Beta mercaptoethanol
  • Concentration information loading...
  • Lysate notes
    BALB/3T3 clone A31 cell lysate was prepared by homogenization in modified RIPA buffer(150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl flouride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 µg/ml of aprotinin, 5 µg/ml of leupeptin). Cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The cell lysate was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol.
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab7901 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. Ready to load; 20 µg/lane is recommended for mini gel.

References

This product has been referenced in:
  • Dara L  et al. Receptor interacting protein kinase 1 mediates murine acetaminophen toxicity independent of the necrosome and not through necroptosis. Hepatology 62:1847-57 (2015). Read more (PubMed: 26077809) »
See 1 Publication for this product

Publishing research using ab7901? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

Q&A
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1-2 of 2 Q&A

Question

Hello,

Yesterday, I used your p19ARF antibody(ab80) for my samples. Also I set NIH3T3 whole cell lysate as a positive control, but it didn’t work. (no band.)

I borrowed this cell line from other lab. After I cultured, I extracted lysate by myself.

I think there is some possibility that my NIH3T3 had any problems. So, I plan to order your cell lysate and check some mouse embryonic fibroblast cell line; ab7179,ab14874,ab7901,ab3874.

I’d like to ask you which is the best as positive control for p19ARF antibody? Or is there no difference among them?

Thank you,

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Answer

Thank you very much for contacting us with your enquiry.

I'm sorry to hear that there were no bands in Western blot using this antibody. I would expect any of the MEF cell lines to work as a positive control, but I suggest using a whole cell lysate (ab14874 is a nuclear fraction) since that is what we have tested with ab80. What kind of samples are you using other than the MEF lysate?

If you'd like to send some more details about your protocol ( how much protein was loaded, dilution of antibody, blocking solution, etc) I'd be happy to take a look and see if we have any suggestions to improve the results.

I look forward to hearing from you. If there's anything else that we can do for you, please let me know and I'll be happy to help.

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Question

I am interested in purchasing a fibroblast whole cell lysate product from you company to test a new procollagen probe my lab developed.

I noticed that you sell several kinds of fibroblast lysate products:ab7179, ab3960, ab3874, ab7901...

I wanna know if these products have a high procollagen (˜150KDa) content. Have you tested that? Or do you know if there have been any existing reports in which these products have been used for procollagen detection? This information is important for me to make the decision.

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Answer

Thank you for contacting Abcam and for your interest in our whole cell lysates.

Regarding these products, they are not tested for protein composition. They are only tested for protein concentration and confirmed to not be degraded.

If you are interested in determining which lysates will contain the protein that you need, I would recommend a search of the literature. Unfortunately, we are not experts in this field and would not want to recommend a product that is not suitable for your experiments.

I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

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

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