RIPA Lysis Buffer (Strong)

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Catalogue No: abx090624
Price: US$174.00
(Size: 100 ml)

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RIPA Lysis Buffer (Strong)

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Datasheet SDS Download IFU
RIPA Lysis Buffer (Strong) is a cell and tissue lysis buffer, commonly used in Western blot and Immunoprecipitation applications. This buffer is most effective for denaturing nuclear and membrane proteins, and lysing tissue samples.

Abbexa's RIPA (Radio Immunoprecipitation Assay) Lysis Buffers are available in different strengths to balance protein extraction with structure and function:
  1. abx090624 - Strong (high detergent concentration)
  2. abx090624 - Medium (lower detergent concentration)
  3. abx090626 - Weak (very low detergent concentration)
Protein concentration following lysis can be determined by our BCA Protein Assay Kits (abx090640, abx090641, abx090642 and abx090643). It is not recommended to use a Bradford Protein Assay Kit or other Kinase assay kits when using this buffer.

Target RIPA Lysis Buffer
Research Area Cell Lysis Buffers
Tested Applications WB, IP
Form Liquid
Storage Store at 4°C.
Shelf Life 12 months.
Buffer Contains 50 mM Tris, pH 7.4, 150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, and inhibitors (sodium orthovanadate, sodium fluoride, EDTA and leupeptin).
Availability Shipped within 5-10 working days.
Note THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC, THERAPEUTIC OR COSMETIC PROCEDURES. NOT FOR HUMAN OR ANIMAL CONSUMPTION.
Directions for use

Cultured Cell Samples

  1. Prepare an appropriate volume of RIPA Lysis Buffer. Add PMSF Buffer (protease inhibitor) approximately 2-3 min before use to make a final PMSF concentration of 1 mM, and mix thoroughly.
    2. Plate Size / Surface AreaRecommended Buffer Volume
    60 mm2250-500 µl
    100 mm2500-1000 µl
    6-well cell culture plate200-400 µl per well
    24-well cell culture plate100-200 µl per well
    96-well cell culture plate50-100 µl per well
  2. For adherent cells, wash the sample with PBS, normal saline or serum-free culture medium to remove the culture solution. Add an appropriate volume of RIPA Lysis Buffer, then mix until the cells immerse completely in the buffer. Shake gently for 5-10 min. After lysis, centrifuge at 10,000-14,000 × g for 10 min, then collect the supernatant.

    For suspension cells, centrifuge the sample to collect the cells, then wash the sample with PBS, normal saline or serum-free culture medium. Add an appropriate volume of RIPA Lysis Buffer, then mix until the cells are dispersed. Vortex for 5-10 min to fully lyse the cells. There should be no visible precipitate after lysis; for large volumes of cells, aliquot then lyse. After lysis, centrifuge at 10,000-14,000 × g for 10 min, then collect the supernatant.

Tissue Samples

  1. Rinse the tissue sample in cold saline to remove any excess blood. Weigh the sample, then cut it into smaller pieces and place in a tissue homogeniser.
  2. Add an appropriate volume of RIPA Lysis Buffer and mix thoroughly. Add PMSF Buffer (protease inhibitor) approximately 2-3 min before use to make a final PMSF concentration of 1 mM.
    Note: It is recommended to use a Net Tissue Weight (g) to RIPA Lysis Buffer (ml) ratio of 1:10 (e.g. add 10 ml RIPA Lysis Buffer to 1 g of tissue). For incompletely lysed tissues, the volume of RIPA Lysis Buffer can be increased. For samples with high protein concentration, the volume of RIPA Lysis Buffer can be reduced.
  3. Homogenise with a glass homogeniser until samples are completely lysed.
  4. Centrifuge at 10,000-14,000 × g for 10 min, then collect the supernatant.

Notes

  • It is normal to see a small transparent jelly substance as an end product after adding RIPA Lysis Buffer. This transparent jelly is a complex containing genomic DNA. If genomic-DNA binding proteins are not of interest, the end user can centrifuge and collect the supernatant as normal. Otherwise, the jelly can be dispersed by ultrasound (e.g. sonication) before centrifugation. Ultrasound is not necessary for detecting transcription factors such as NF-kappa B and p53.
  • All steps of the protein extraction should be performed at 4 °C or on ice. It is recommended to aliquot large samples into smaller volumes, and then lyophilizing or storing at -20 °C in liquid form. Avoid repeated freeze/thaw cycles.
  • Personal protective equipment (PPE), such as lab coats, lab glasses and disposable gloves, are recommended when using this product.
Research Articles on RIPA Lysis Buffer


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