NIH 3T3 whole cell lysate (ab7179)

Overview

  • Product nameNIH 3T3 whole cell lysate
    See all NIH 3T3 lysates
  • General notesCell line: NIH/3T3 (Mouse embryonic fibroblast).
    Growth media: DMEM and 10% bovine calf serum.


    Mouse NIH/3T3 cell lysate was prepared by homogenization in modified RIPA buffer(50 mM Tris-HCl, pH 7.4, 1% Triton X-100, 0.2% sodium deoxycholate, 0.2% sodium dodecylsulfate (SDS), 1 mM sodium ethylenediaminetetraacetate,1 mM phenylmethyl-sulfonyl flouride, 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 lysate was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% SDS, 0.01% bromophenol blue) containing 5% b-mercaptoethanol.

  • Tested applicationsSuitable for: WBmore details

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
  • Storage bufferpH: 6.80
    Constituent: 5% Beta mercaptoethanol
  • Concentration information loading...
  • PurityWhole Cell Lysate
  • Lysate notesMouse NIH/3T3 cell lysate was prepared by homogenization in modified RIPA buffer(50 mM Tris-HCl, pH 7.4, 1% Triton X-100, 0.2% sodium deoxycholate, 0.2% sodium dodecylsulfate (SDS), 1 mM sodium ethylenediaminetetraacetate,1 mM phenylmethyl-sulfonyl flouride, 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 lysate was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% SDS, 0.01% bromophenol blue) containing 5% b-mercaptoethanol.
  • Research areas
  • BackgroundNIH 3T3 cells are established from a NIH Swiss mouse embryo. These cells are highly contact inhibited and are sensitive to sarcoma virus focus formation and leukaemia virus propagation. Cells have now lost their contact inhibition. This cell line was established from NIH Swiss mouse embryo cultures in the same manner as the original random bred 3T3 and the inbred BALB/c 3T3. The established NIH/3T3 line was subjected to more than 5 serial cycles of subcloning in order to develop a subclone with morphologic characteristics best suited for transformation assays. It is therefore used for DNA transfection studies

Applications

Our Abpromise guarantee covers the use of ab7179 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.
NIH/3T3 cell lysate is ready to load on SDS-PAGE for Western blotting, 20 µg per lane is recommended for mini gel.

References for NIH 3T3 whole cell lysate (ab7179)

ab7179 has not yet been referenced specifically in any publications.

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Thank you for contacting us.

Based on this image alone it seems like the lower band is your CDKN2A/p19ARF band. It is possible for band size to change slightly due to sample preparation or buffer, however usually the positive control will be...

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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 ...

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I apologize for the delay in getting back to you. I am still waiting for a reply from our source of the skeletal muscle lysate. I do not think it should contain alpha-SMA.

On the other hand, the fault may be with the antibody, if is not dete...

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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.
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Thank you for your enquiry. You are correct, this antibody is at a concentration of 2mgml-1. The antibody is provided at 100 µg therefore 50ul. We do receive some very occasional shortages which requires our lab team taking a vial and adding lysate...

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