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NIH 3T3 (Mouse embryonic fibroblast cell line) Nuclear lysate (ab14874)

Q&A (6)

Overview

  • Product nameNIH 3T3 (Mouse embryonic fibroblast cell line) Nuclear lysateSee all NIH 3T3 lysates ...
  • General notesExtracts have been quality control tested by Western blot and the Electrophoretic Mobility Shift Assay (EMSA).

Properties

  • FormLiquid
  • Storage instructionsAliquot and store at -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: None
    Constituents: 20% Glycerol, 20mM HEPES, 100mM Potassium Chloride, 1mM Magnesium Chloride, 0.5mM PMSF, 0.5mM DTT. pH 7.9
  • Concentration information loading...
  • 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

    • References for NIH 3T3 (Mouse embryonic fibroblast cell line) Nuclear lysate (ab14874)

    ab14874 has not yet been referenced specifically in any publications.

    Publishing research using ab14874 ? Please let us know so that we can cite the reference in this datasheet

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    Displaying 1 - 6 of 6 results for Q&A

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    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 extract...

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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 lys...

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    Thanks very much for your prompt reply and for checking for possible related proteins. For your questions:

    How much lysate per lane? 30 ug in the first lane, 15 ug in the second (indicated at the top of the blot)

    Blocking solution? M...

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    Thank you very much for your reply and for the additional information.

    Have you tried any other exposure times? Since the specific band seems to be the strongest (with 1:100 of antibody and 15-30 ug of protein), you can probably reduce the expos...

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    Hi--I recently ran a Western with OGG1 antibody ab91421 and 3T3 nuclear lysate ab14874. I got the bands near 39 & 60kD as shown in

    http://www.abcam.com/Ogg1-antibody-ab91421.html

    but also many other bands (below). Do you have sug...

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    Thank you very much for contacting us with your question and for sending the image of your results.

    According to the UniProt page for this protein, there aren't any known post-translational modifications like cleavage or glycosylations that migh...

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    Do you have any Ezh1 antibody that works well on mouse tissue?

    Thank you for contacting us. We have three antibodies that may be suitable for your purposes:

    ab13665: rabbit polyclonal to EZH1 validated in WB on mouse and human samples
    ab86128: rabbit polyclonal to EZH1 validated in WB and ICC/IF on mou...

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    I ran a western blot with your cell lysate and blotted with Ezh1 Ab at
    1:5000 and then Donkey anti-RHRP 1:100,000 dilution. The whole blot turned
    black after 1 s exposure time.
    I tried to figure out which reagent causing the high background...

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    Thank you for sending the image of your blots.

    When you use the PVDF membrane, are you pre-wetting it with methanol? Because PVDF is hydrophobic, it may not properly bind the blocking agent without the pre-wetting step. This could potentially ca...

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    My WB membrane is completely black using ab64850.

    Thank you for your phone call. As we discussed, it is possible that your 6 x 15 minute washes are washing the blocking agent off your membrane. It may help to reduce the wash time to 3 x 5 min, shorten the primary antibody incubation to 1 hour at RT, and ...

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