NP-40 Lysis Buffer

Item Number

N748603

• Product Name: NP-40 Lysis Buffer

Product Description

Aladdin's NP-40 Lysis Buffer is a relatively mild lysis buffer for cells and tissues. The obtained sample lysate can be used for routine PAGE, Western, IP, Co-IP, and ELISA analysis.This product can be used to lyse animal, plant, bacteria and yeast samples.For the main characteristics and comparison of Aladdin's lysis buffers, please visit the website at https

Precautions：

For the best performance, this product should be avoided repeated freeze-thaw. Store aliquots at -20℃.PMSF (, ST506) is required, but not supplied in this product. Inhibitor cocktails can also be used for better lysis results. provides a variety of inhibitor cocktails, such as Protease and Phosphatase Inhibitor Cocktail for General Use , Protease and Phosphatase Inhibitor Cocktail for General Use, MS-safe , Protease and Phosphatase Inhibitor Cocktail for Mammals , Protease and Phosphatase Inhibitor Cocktail for plant , Protease and Phosphatase Inhibitor Cocktail for Fungi , and Protease and Phosphatase Inhibitor Cocktail for Bacteria . If phosphorylated proteins are not to be detected, inhibitors of protease only can be used.All procedures for sample lysis should be performed on ice or at 4℃.An appropriate sample lysis buffer can be selected by referring to the website at https://www.aladdin-e.com or optimized by preliminary tests.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.

Instructions for Use：

1. Lysis of cultured cellsa. Thaw Lysis Buffer completely and mix well. Just before use, take an appropriate amount of Lysis Buffer and add PMSF to a final concentration of 1mM. Other protease and phosphatase inhibitors can also be used if required.b. For adherent cells: Aspirate the culture medium and wash cells once with PBS, buffered saline or serum-free culture medium to remove protein residues that may interfere with cell lysis. Add 150-250μl of Lysis Buffer for each well of cells in 6-well plates and pipette up and down several times to ensure full contact of cells with the Lysis Buffer. Generally, it takes 1-2 sec to lyse animal cells or 2-10 min to lyse plant cells.For suspension cells: Collect cells by centrifugation, vortex gently or flick the bottom of the tube to disperse cells as much as possible. Add 150-250μl of Lysis Buffer for each well of cells in 6-well plates, flick the bottom of the tube to resuspend and lyse cells thoroughly. There should be no obvious precipitates after full lysis. For large amounts of cells, dispense them into 0.5-1.0×106 cells per tube for lysis. For bacteria or yeasts: Collect bacteria or yeasts by centrifuging 1ml of cultures. Aspirate the supernatant and wash the pellet once with PBS if necessary. After removing the supernatant, vortex gently or flick the bottom of the tube to disperse cells, and then add 150-250μl of Lysis Buffer. After resuspending cells completely in the Lysis Buffer, lyse cells on ice for 2-10 min. For better lysis, bacteria or yeasts can be treated with lysozyme or lyticase before adding the Lysis Buffer.Note: Generally, for one well of cells in 6-well plates or 1ml of bacterial or yeast cultures, 150μl of Lysis Buffer is sufficient. For higher cell densities, use 200-250μl of Lysis Buffer. When lysing 1 million animal cells with 100μl of Lysis Buffer, the protein concentration of the lysate is approximately 2-4 mg/ml, which is dependent on the type of cells. c. After full lysis of cells, centrifuge at 10,000-14,000×g for 3-5 minutes and take the supernatant for PAGE, Western or IP analysis. 2. Lysis of tissuesa. Cut tissue into small pieces.b. Thaw the Lysis Buffer completely and mix well. Just before use, take an appropriate amount of Lysis Buffer and add PMSF to a final concentration of 1mM. Other protease and phosphatase inhibitors can also be used if required.c. Add 150-250μl of Lysis Buffer per 20mg of tissues. The amount of Lysis Buffer can be adjusted based on the lysis results.d. Homogenize tissues thoroughly with a glass homogenizer or 's TissueMasterTM Handheld Homogenizer . e. Centrifuge at 10,000-14,000×g for 3-5 minutes and take the supernatant for PAGE, Western or IP analysis. When lysing 20mg of mouse liver tissue with 200μl of Lysis Buffer, the protein concentration of the lysate is approximately 15-25 mg/ml, which is dependent on the state of tissues.f. For tiny tissues, add Lysis Buffer after appropriate cutting and lyse tissues with vigorous vortex. Centrifuge and take the supernatant for subsequent analysis. This lysis method is convenient, with no homogenization needed, but the lysis result is not as thorough as that from the homogenization method.Appendix：Comparison and selection of Lysis Buffers from : Cat. No. Component QuantityP0013P0013BP0013CP0013DP0013FP0013GP0013JP0013KProduct NameLysis Buffer for Western&IPRIPA Lysis Buffer RIPA Lysis Buffer (Medium)RIPA Lysis Buffer (Weak)NP-40 Lysis BufferSDS Lysis BufferLysis Buffer for Western&IP (No Inhibitors)RIPA Lysis Buffer Effective Component1% Triton X-1001% Triton X-100, 1% deoxycholate, 0.1% SDS1% NP-40, 0.5% deoxycholate, 0.1% SDS1% NP-40, 0.5% deoxycholate 1% NP-401% SDS1% Triton X-1001% Triton X-100, 1% deoxycholate, 0.1% SDSLysis StrengthMildStrongMediumMildMildStrongMildStrongExtraction of Membrane ProteinsFairExcellentGoodFairFairExcellentFairExcellentExtraction of Cytosolic ProteinsExcellentExcellentExcellentExcellentExcellentExcellentExcellentExcellentExtraction of Nuclear ProteinsGoodExcellentGoodGoodGoodExcellentGoodExcellentExtraction of Phosphorylated Proteins in CytosolExcellentExcellentExcellentExcellentExcellentExcellentExcellentExcellentExtraction of Transcription Factors in NucleusExcellentExcellentExcellentExcellentExcellentExcellentExcellentExcellentProtease InhibitorsYesYesYesYesYesYesNoNoPhosphatase InhibitorsYesYesYesYesYesYesNoNoSpecies CompatibilityHighHighHighHighHighHighHighHighApplicationsWB, IP, Co-IPWB, IPWB, IPWB, IP, Co-IPWB, IP, Co-IPWB, ChIPWB, IP, Co-IPWB, IPFor general Western, IP or co-IP assays, we recommend using Lysis Buffer for Western & IP (, P0013) which has been extensively used with excellent performance. Lysing cells with this Lysis Buffer does not need ultrasonic processing. The obtained sample lysate has no gelatinous DNA clump and is ideal for subsequent studies as well as detection of phosphorylated proteins by Western.For IP of some special proteins, RIPA Lysis Buffer or NP-40 Lysis Buffer can be attempted when Lysis Buffer for Western & IP (, P0013) does not work well. If the background of IP result is high, a stronger Lysis Buffer such as RIPA Lysis Buffer should be used. If the target protein can not be immunoprecipitated, it indicates that the Lysis Buffer used is too strong, and a milder Lysis Buffer such as RIPA Lysis Buffer (Weak) and NP-40 Lysis Buffer should be tried.For Western blot of some proteins hard to be soluble, RIPA Lysis Buffer or SDS Lysis Buffer can be attempted when Lysis Buffer for Western & IP (, P0013) does not work well. RIPA Lysis Buffer is also extensively used by investigators and has been cited in large amounts of articles.For special applications where no inhibitors or a particular inhibitor is needed, P0013J or P0013K can be selected. P0013J is compatible with enzymatic activity assay and small molecule detection in most cases, but for special enzymes or small molecules, users should check the compatibility on their own. P0013J has weaker lysis capability than P0013K, but P0013J generally has better compatibility with enzyme and small molecule studies.