N748603A: Store at 2-8℃ long term (12 months). Upon receipt, it is recommended to aliquot. N748603B & N748603C Store at -20°C long term (12 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.
Storage Temp
Store at 2-8°C,Store at -20°C,Avoid repeated freezing and thawing
Shipped In
Ice chest + Ice pads
This product requires cold chain shipping. Ground and other economy services are not available.
Grade
Bioac(废弃), BioReagent, for western blot, 废弃PharmPure™ Apex Ph Eur
Product Description
N748603
Component
100mL
500mL
Storage
N 748603A
NP-40 cracking solution
100mL
500mL
2-8℃
N748603B
50x phosphatase inhibitor
2x1mL
10x1mL
-20℃. Avoid freeze/thaw cycle.
N748603C
100xPMSF
1mL
5x1mL
-20℃. Avoid freeze/thaw cycle.
NP-40 Lysis Buffer is a relatively mild cell tissue lysis buffer, and the protein samples obtained from its lysis can be used for conventional Western, IP, and co IP assays. The protein sample obtained by lysing with NP-40 lysate can be measured for protein concentration using the BCA protein concentration assay kit. Due to the high concentration of descaling agents, the Bradford method cannot be used to determine the protein concentration of the sample obtained from the lysis of this lysate. Precautions： 1. To achieve the best usage effect, it can be appropriately packaged and stored at -20 ℃ to avoid excessive repeated freezing and thawing; 2. All steps of sample cracking must be carried out on ice or at 2-8 ℃; 3. For your own safety, please take protective measures such as wearing lab coats and gloves before using the reagents. Instructions for Use： 1. Reagent preparation Take an appropriate amount of lysis buffer and add the protease inhibitor complex and/or phosphatase inhibitor complex to the lysis buffer at a ratio of 1:50 within a few minutes before use, or use 100 mM PMSF and make the final concentration of PMSF 1 mM. 2. Cell/tissue lysis For adherent cells: Remove the culture medium and wash once with PBS, physiological saline, or serum-free culture medium. Add lysis buffer in a ratio of 150-250 uL per well to a 6-well plate. Blow with a gun several times to ensure full contact between the lysate and the cells. Usually, after 1-2 seconds of contact with the cell lysate, the cell will be lysed. If used for ChIP, it needs to be further cracked on an ice bath for 10 minutes after initial cracking; For suspended cells: Collect cells by centrifugation and use your fingers to forcefully scatter them. Add lysis buffer at a ratio of 150-250uL per well of cells in a 6-well plate. Gently flick with your fingers to fully lyse the cells. If there are a large number of cells, they must be divided into 500000 to 1 million cells/tube and then lysed. After sufficient lysis, there should be no obvious cell precipitation. If used for ChIP, it needs to be further cracked on an ice bath for 10 minutes after initial cracking. For tissue samples: Cut the tissue into fragments using a tissue cutter, add 150-250uL of lysis buffer per 20mg of tissue, and homogenize with a glass homogenizer until fully lysed. If used for ChIP, it needs to be further cracked on an ice bath for 10 minutes after initial cracking. 3. Collection of protein samples After sufficient lysis, centrifuge at 10000-14000g for 3-5 minutes, take the supernatant, and proceed with subsequent PAGE, Western blot, and ChIP operations.
