Product Information
Product Usage Information
1. If buffer will be used regularly, it is recommended that the 1X buffer be kept at 4ºC for 1-2 weeks. For longer periods of time, buffer should be stored at -20ºC. Aliquotting of 1X buffer is recommended to avoid freeze thaw cycles.
2. Thaw 1X buffer at 24-30ºC, mixing end-over-end.
3. Chill 1X buffer on ice and add PMSF just prior to use.
Note: Cell Signaling Technology recommends adding 1 mM PMSF (final concentration) immediately before use.
Lysis:
For lysis of adherent cells, we recommend the following: (all reagents and lysates must be kept cold)
1. Treat cells as desired.
2. Wash plate with ice-cold PBS to remove residual media.
3. Add 500–1000 µl of lysis buffer/ 10 cm dish.
4. Incubate plate on ice for 2 minutes.
5. Collect cell lysate solution for use. Spin extract 1 minute at 14,000 x g in a cold microfuge if cell debris is present.
Additional notes:
1. For non-adherent cells, judgment must be used as to amount of lysis buffer to add. Addition of volume equal to cell pellet is generally appropriate.
2. Additional protease inhibitors can be added to the 1X lysis buffer if needed.
Solutions and Reagents
1X Lysis Buffer: 20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM Na2 EDTA, 1 mM EGTA, 1% Triton, 20 mM sodium Pyrophosphate, 25 mM Sodium Fluoride, 1 mM β-glycerophosphate, 1 mM Na3VO4, 1 µg/ml leupeptinNote: Cell Signaling Technology recommends adding 1 mM PMSF immediately before use.
Storage
Product Description
Background
PathScan® Sandwich ELISA Lysis buffer (1X) was developed for ELISA applications and is formulated to better inhibit Serine / Threonine class of phosphatases after cell extracts are made.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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