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Product Datasheet

10X Lysis Buffer

Overview
Catalog #100-LYS

Size

15 mLFormat

SDS Lysis Buffer

Details
Product Description

10X Lysis Buffer optimized to fully solubilize proteins in preparation for western blotting. May be
used for organs and tissue culture cells.

Background

Nonionic detergents such as NP-40 or triton fail to solubilize many cellular proteins involved in cellsignaling. This problem is particularly acute in brain where synaptic junctions are known to be
insoluble in nonionic detergents. 1% SDS completely solubilizes membrane and other hard tosolubilize proteins including synaptic junction proteins. Suitable for both organs and tissue culturecells.
•SDS inactivates most proteases, but additional protease inhibitors can be added as needed
before use as some proteases are insensitive to or even activated (e.g. proteinase K) bySDS.

•

1% SDS inactivates phosphatase enzymes. If treating your lysate with phosphatase
enzymes this lysis buffer is not recommended.

Buffer10% (w/v) SDS, 100 mM TRIS, 10 mM EDTA, pH 8.0. Dilute to 1X with dH2O before use.

Storage

Storage at room temperature is recommended for both 10X and 1X solutions to prevent SDS fromprecipitating out of solution.

Lysate Preparation Protocol using 1X Lysis Buffer
Prepare Samples

A. Organs- Whole cell lysate
1. Place organ in conical tube or microfuge tube with screw cap. For large organs cut into
1/4”sections. For organs that are thick and dense cut even smaller sections.
2. Add lysis buffer to organs in conical tube completely submerging the organ in the lysis
buffer. Keep sample at room temperature.

B. Adherent Tissue Culture Cells
1. Wash cells with 2-10 mLs of room temperature 1X PBS once. Remove wash with transferpipet.
2. Add lysis buffer, enough to cover the entire plate.
3. Rock and rotate the plate to thoroughly coat cells.
4. Transfer to a conical tube or microfuge tube with screwcap.
C. Suspension TC Cells
1. Pipette media and cells into conical tube.
2. Pellet cells by centrifugation (1 minute at 1800 x g) and remove media.
3. Resuspend cells with 1-3 mLs of room temperature 1X PBS to wash.
4. Repeat step #2.
5. Add lysis buffer.
Lyse Samples

1.
2.
3.
4.

Sonicate sample in 5-20 second intervals until buffer is clear and can be easily pipetted
without clogging.
Heat sample at 95°C for 10 minutes. Cool to room temperature.
Centrifuge lysate at 1800 x g for 5 minutes to pellet cell debris.
Pipette lysate into fresh conical or microfuge tube.

Significant Citations
Brett, J.O., Dubash, T.D., Johnson, G.N., Niemierko, A., Mariotti, V., Kim, L.S., Xi, J., Pandey, A., Dunne, S., Nasrazadani, A. and Lloyd, M.R., 2023. A
Gene panel associated with abemaciclib utility in ESR1-mutated breast cancer after prior cyclin-dependent kinase 4/6-inhibitor progression. JCO
Precision Oncology, 7, p.e2200532.
Palliyage, G.H., Samart, P., Bobbala, S., Rojanasakul, L.W., Coyle, J., Martin, K., Callery, P.S. and Rojanasakul, Y., 2023. Chemotherapy-induced PDL-1
expression in cancer-associated fibroblasts promotes chemoresistance in NSCLC. Lung Cancer, 181, p.107258.
Endo, Y., Hwang, C.D., Zhang, Y., Olumi, S., Koh, D.J., Zhu, C., Neppl, R.L., Agarwal, S. and Sinha, I., 2023. VEGFA Promotes Skeletal Muscle
Regeneration in Aging. Advanced Biology, p.2200320.
Jablon, K.L., Akerstrom, V.L., Li, M., Braun, S.E., Norton, C.E. and Castorena‐Gonzalez, J.A., 2022. Isolation and Short‐Term Culturing of Primary
Lymphatic Endothelial Cells from Collecting Lymphatics: A Techniques Study. Microcirculation, p.e12778.
Lloyd, J.T., Yee, A.G., Kalligappa, P.K., Jabed, A., Cheung, P.Y., Todd, K.L., Karunasinghe, R.N., Vlajkovic, S.M., Freestone, P.S. and Lipski, J., 2022.
Dopamine dysregulation and altered responses to drugs affecting dopaminergic transmission in a new dopamine transporter knockout (DAT-KO) rat
model. Neuroscience, 491, pp.43-64.
Cerqueira, S.R., Benavides, S., Lee, H.E., Ayad, N.G. and Lee, J.K., 2022. BET protein inhibition promotes non-myeloid cell mediated neuroprotection
after rodent spinal cord contusion. Experimental Neurology, 352, p.114035.

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