Cell and Tissue Lysis Buffer for Nitric Oxide Assay

Cat. No.: C752147

Overview

Precautions：

All procedures should be performed on ice or at 4℃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. Preparation of cell samples a. Thaw Cell and Tissue Lysis Buffer for Nitric Oxide Assay and mix well before use.b. For adherent cells: Remove the culture medium and wash the cells once with PBS, saline, or serum-free medium. Add 100-200μl of this product to each well of a 6-well plate and pipette several times to ensure sufficient contact of cells with the lysis buffer. Cells are typically lysed after 1-2 seconds.c. For suspension cells: Collect cells by centrifugation, vortex gently or flick the bottom of the tube to disperse cells. Add 100-200μl of this product to each well of cells in 6-well plates, flick the bottom of the tube to resuspend and lyse cells thoroughly. For large amounts of cells, dispense them into 0.5-1.0×106 cells per tube for lysis.d. After complete lysis, centrifuge at 10,000-14,000×g for 3-5 minutes, and take the supernatant for NO assay or protein concentration determination. If the NO assay cannot be completed on the same day after sample preparation, store the supernatant at -20℃ and complete the assay as soon as possible.Note: Usually, 100μl of this product for each well of cells in 6-well plates is sufficient, but if the cell density is very high, the amount of lysis buffer can be increased to 150μl or 200μl.2. Preparation of tissue samples a. Thaw Cell and Tissue Lysis Buffer for Nitric Oxide Assay and mix well before use.b. Cut the tissue into small pieces.c. Add 100-200μl of this product per 20mg of tissue. The amount of lysis buffer can be adjusted based on the lysis results.d. Homogenize tissues thoroughly with the TissueMasterTM Handheld Homogenizer (, E6600) or other suitable homogenization devices. For tiny tissues, add lysis buffer after appropriate cutting and lyse 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.e. After complete lysis, centrifuge at 10,000-14,000×g for 3-5 minutes, and take the supernatant for NO assay or protein concentration determination. If the NO assay cannot be completed on the same day after sample preparation, store the supernatant at -20℃ and complete the assay as soon as possible.

Specifications

Product Name

Cell and Tissue Lysis Buffer for Nitric Oxide Assay

Molecule Type

Bioreagents/Buffer

Storage and Shipping

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Stability And Storage

Store at -20℃ for up to 1 year.