Cell Lysis Buffer

This Cell Lysis Buffer is a ready-to-use Western blot related reagent solution used for efficient extraction of total soluble protein in nondenatured state from mammalian cells. Cell lysis can be finished in 30 minutes. The cell lysis buffer has been validated for use with COS-7, NIH3T3, Hepa 1-6, 293T, CHO and other cell types. It is able to lyse plated cells and pelleted cells from suspension mammalian cultures. The produced cell lysates are directly compatible with reporter gene expression assays (luciferase, beta-galactosidase, chloramphenicol acetyl transferase, CAT, alkaline phosphatase), protein kinase assays (PKA, PKC, tyrosine kinase), phosphatase assays (general phosphatases, tyrosine phosphatases), immunoassays (Western blots, ELISAs, RIAs, immunoprecipitation), Coomassie-Blue and silver staining, BCA protein assay, protein purification procedures, electrophoresis, and many other downstream applications. Cited in 24 publication(s).

Product Info Summary

SKU AR0103
Pack size 100 mL (The kit contains sufficient lysis buffer for 200 cell pellet fractions containing 5x 106 cells each)
Applications Immunoassays: WB, ELISA, RIA, immunoprecipitation; reporter assays; protein kinase assays; protein purifications

Overview

Physical State Liquid
Pack Size 100 mL (The kit contains sufficient lysis buffer for 200 cell pellet fractions containing 5x 106 cells each)
Description Boster’s Cell Lysis Buffer is a ready-to-use Western blot related reagent solution used for efficient extraction of total soluble protein in nondenatured state from mammalian cells.
Storage & Expiration Upon receipt store at 4°C. Cell Lysis Buffer is stable for one year. Product is shipped on ice.
Equivalent Thermofisher (Product No. 78501, 78503, 78505); Millipore Sigma (Product No. C2978)
Application Immunoassays: WB, ELISA, RIA, immunoprecipitation; reporter assays; protein kinase assays; protein purifications
*Our Boster Guarantee covers the use of this product in the above tested applications.
Enzymatic Activity None; use additives to add function
Additional components compatibility Compatible with additives, e.g. protease inhibitors (PMSF), reducing agents, chelating agents, salts
Cite This Product Cell Lysis Buffer (Boster Biological Technology, Pleasanton CA, USA, Catalog # AR0103)

Description

This Cell Lysis Buffer is a ready-to-use Western blot related reagent solution used for efficient extraction of total soluble protein in nondenatured state from mammalian cells. Cell lysis can be finished in 30 minutes. The cell lysis buffer has been validated for use with COS-7, NIH3T3, Hepa 1-6, 293T, CHO and other cell types. It is able to lyse plated cells and pelleted cells from suspension mammalian cultures. The produced cell lysates are directly compatible with reporter gene expression assays (luciferase, beta-galactosidase, chloramphenicol acetyl transferase, CAT, alkaline phosphatase), protein kinase assays (PKA, PKC, tyrosine kinase), phosphatase assays (general phosphatases, tyrosine phosphatases), immunoassays (Western blots, ELISAs, RIAs, immunoprecipitation), Coomassie-Blue and silver staining, BCA protein assay, protein purification procedures, electrophoresis, and many other downstream applications.

Additional Materials Required

• Protease inhibitor (Product No. AR1182) and phosphatase inhibitor (Product No. AR1183)

• 2 ml microcentrifuge tubes

• Microcentrifuge capable of spinning at 10,000 x g

• Cell Scraper

Procedure for Lysis of Monolayer-cultured Adherent Mammalian Cells

Note: Pre-chill an appropriate volume of Cell Lysis Buffer at 4°C. If desired, add protease inhibitor and phosphatase inhibitor to the lysis buffer immediately before use.

1. In a microcentrifuge tube, resuspend 5×106 cells in the growth media by scraping the cells off the surface of the plate with a cell scraper. Centrifuge harvested cell suspension at 600xg for 5min, then carefully remove and discard the supernatant.

2. Resuspend the cells in chilled PBS. Centrifuge at 600xg for 5min, then carefully remove and discard the supernatant.

3. Add 0.5 mL of chilled cell lysis buffer to the cell pellet. Vortex briefly. Incubate on ice for 30 minutes.

4. Centrifuge samples at 10000xg for 10 minutes.

5. Transfer supernatant to a new tube for further analysis.


Note: Cell lysis buffer can be added directly to the flask containing cells. Please see the following procedures.

1. Carefully remove culture medium from adherent cells.

2. Wash cells with chilled PBS. Carefully remove PBS.

3. Add chilled Cell lysis buffer to the cells. Vortex briefly. Incubate on ice for 30 minutes. (For the volume of the lysis buffer, follow the instructions listed below)

SIZE of the plate/surface areaVolume of the lysis buffer
100mm500-1000μL
60mm250-500μL
6-well plate200-400μL per well
24-well plate100-200μL per well
96-well plate50-100μL per well

4. Centrifuge samples at 10000xg for 10 minutes.

5. Transfer supernatant to a new tube for further analysis.

Procedure for Lysis of Suspension-cultured Mammalian Cells

Note: Pre-chill an appropriate volume of Cell Lysis Buffer at 4°C. If desired, add protease inhibitor and phosphatase inhibitor to the lysis buffer immediately before use.

1. In a microcentrifuge tube, harvest 5×106 cells by centrifugation at 600xg for 5min. Carefully remove and discard the supernatant.

2. Resuspend the cells in chilled PBS. Centrifuge at 600xg for 5min, then carefully remove and discard the supernatant.

3. Add 0.5 mL of chilled Cell lysis buffer to the cell pellet. Vortex briefly. Incubate on ice for 30 minutes.

4. Centrifuge samples at 10000xg for 10 minutes.

5. Transfer supernatant to a new tube for further analysis.

Precautions

• All steps of protein lysis should be operated on ice or at 4 ˚C.

• Use BCA Protein Assay kit (Product No. AR0146) to quantify lysed proteins. Bradford Protein Assay kit is not recommended.

Product Images

AR0103 has been cited in 24 publications:

*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.

Huang, Y., Zhang, J., Xu, D., Peng, Y., Jin, Y., & Zhang, L. (2021). SIRT6‑specific inhibitor OSS‑128167 exacerbates diabetic cardiomyopathy by aggravating inflammation and oxidative stress. Molecular Medicine Reports, 23, 367. https://doi.org/10.3892/mmr.2021.12006
Species: Mouse,Rat
AR0103 usage in article: APP:WB, SAMPLE:H9C2 CELL, DILUTION:NA

Pedicini L,Wiktor SD,Simmons KJ,Money A,McKeown L.Affinity-based proteomics reveals novel binding partners for Rab46 in endothelial cells.Sci Rep.2021 Feb 18;11(1) :4054.doi:10.1038/s41598-021-83560-y.PMID:33603063;PMCID:PMC7893075.
Species: Human
AR0103 usage in article: APP:WB, SAMPLE:COS-7 CELL, DILUTION:NA

Ge W,Wang T,Zhao Y,Yang Y,Sun Q,Yang X,Gao Y,Xu X,Zhang J.Period1 mediates rhythmic metabolism of toxins by interacting with CYP2E1.Cell Death Dis.2021 Jan 12;12(1):76.doi:10.1038/s41419-020-03343-7.PMID:33436540;PMCID:PMC7804260.
Species: Mouse
AR0103 usage in article: APP:IP, SAMPLE:LIVER TISSUE, DILUTION:NA

Qi Xiong,Xiang Tian,Weiling Li,Lin Chen,Mei Zhou,Congyue Xu,Qin Ru Sulforaphane alleviates methamphetamine-induced oxidative damage and apoptosis via the Nrf2-mediated pathway in vitro and in vivo.Food and Agricultural Immunology31(1)2020.https://doi.org
Species: Rat
AR0103 usage in article: APP:WB, SAMPLE:PC12 CELLS, DILUTION:NA

Liu L,Dai B,Li R,Liu Z,Zhang Z.Intravital molecular imaging reveals the restrained capacity of CTLs in the killing of tumor cells in the liver.Theranostics.2021 Jan 1;11(1):194-208.doi:10.7150/thno.44979. PMID:33391470;PMCID:PMC7681101.
Species: Mouse
AR0103 usage in article: APP:WB, SAMPLE:LIVER AND SPLEEN, DILUTION:NA

Effect of Chaihushugan San on expression of the Raf/mitogen-activated protein kinase/extracellular signal-regulated kinase pathway in the hippocampi of perimenopausal rats induced by immobilization stress.

Neuroprotective effect of chondroitin sulfate on SH-SY5Y cells overexpressing wild-type or A53T mutant ?-synuclein

Silencing DNA methyltransferase 1 leads to the activation of the esophageal suppressor gene p16 in vitro and in vivo

The Central Role of IFI204 in IFN-? Release and Autophagy Activation during Mycobacterium bovis Infection

Cigarette smoke extract promotes proliferation of airway smooth muscle cells through suppressing C/EBP-? expression

Hypoxia inducible factor 1? promotes survival of mesenchymal stem cells under hypoxia

Blood-filled cerebrospinal fluid-enhanced pericyte microvasculature contraction in rat retina: A novel?in vitro?study of subarachnoid hemorrhage

AIM2 inhibits?autophagy?and IFN-? production during M bovis infection

Chemosensitivity enhancement toward arsenic trioxide by inhibition of histone deacetylase in NB4 cell line

MicroRNA-29b inhibits TGF-?1-induced fibrosis via regulation of the TGF-?1/Smad pathway in primary human endometrial stromal cells

?-catenin knockdown inhibits the proliferation of human glioma cells?in vitro?and?in vivo

Wang Q, Jiang S, Song A, Hou S, Wu Q, Qi L, Gao X. Onco Targets Ther. 2017 Sep 28;10:4753-4763. doi: 10.2147/OTT.S143787. eCollection 2017. HOXD-AS1 functions as an oncogenic ceRNA to promote NSCLC cell progression by sequestering miR-147a

Yu J, Li L, Huang C. Med Sci Monit. 2017 Nov 21;23:5542-5549. Downregulation of Inhibition of Apoptosis-Stimulating Protein of p53 (iASPP) Suppresses Cisplatin-Resistant Gastric Carcinoma In Vitro

Li C, Liu J, Zhang Q, Cui K, Ge Q, Wang C, Chen Z. Int J Oncol. 2018 Jun;52(6):1815-1826. doi: 10.3892/ijo.2018.4346. Epub 2018 Mar 29. Upregulation of E-cadherin expression mediated by a novel dsRNA suppresses the growth and metastasis of bladder...

Piao X, Liu B, Guo L, Meng F, Gao L. Oxid Med Cell Longev. 2017;2017:7085709. doi: 10.1155/2017/7085709. Epub 2017 Jun 21. Picroside II Shows Protective Functions for Severe Acute Pancreatitis in Rats by Preventing NF-κB-Dependent Autophagy

Hu S, Chen Q, Lin T, Hong W, Wu W, Wu M, Du X, Jin R. Oncol Lett. 2018 May;15(5):6171-6178. doi: 10.3892/ol.2018.8118. Epub 2018 Feb 26. The function of Notch1 intracellular domain in the differentiation of gastric cancer

Liu CL, Deng ZY, Du ER, Xu CS. Mol Med Rep. 2018 Apr;17(4):5851-5859. doi: 10.3892/mmr.2018.8601. Epub 2018 Feb 13. Long non-coding RNA BC168687 small interfering RNA reduces high glucose and high free fatty acid-induced expression of P2X7 recepto...

You S, Qian J, Sun C, Zhang H, Ye S, Chen T, Xu Z, Wang J, Huang W, Liang G. J Cell Mol Med. 2018 Mar;22(3):1931-1943. doi: 10.1111/jcmm.13477. Epub 2018 Jan 12. An Aza resveratrol

Chen, C., Cao, M., Wu, D., Li, N., Peng, J., Song, L.,…, & Zhao, J. (2017). KH-type Splicing Regulatory Protein Mediate Inflammatory Response in Gastric Epithelial Cells Induced by Lipopolysaccharide. Cell Biology International. Advance online pub...

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