NIH/3T3 whole cell lysate (ab7179)
Overview
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Product name
NIH/3T3 whole cell lysate
See all NIH 3T3 lysates -
General notes
Cell 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.
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
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Mycoplasma free
Yes -
Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Constituents: 60.05% Water, 12.5% Glycerol (glycerin, glycerine), 9% Tris HCl, 7.7% DTT, 4.4% Sodium chloride, 1% Triton-X-100, 1% Sodium deoxycholate, 1.1% Sodium lauryl sulfate, 0.15% EDTA disodium salt, 0.5% Aprotinin, 0.5% Leupeptin hemisulfate, 0.09% PMSF, 0.01% Bromophenol blue -
Concentration information loading... -
Lysate notes
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. -
Research areas
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Background
NIH 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
The Abpromise guarantee
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. |
| Notes |
|---|
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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. |
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.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab7179 has not yet been referenced specifically in any publications.