NIH/3T3 whole cell lysate (ab7179)
Overview
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Product name
NIH/3T3 whole cell lysate
See all NIH 3T3 lysates -
Description
NIH/3T3 cell lysate -
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.
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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
