Flow cytometric analysis of live mouse splenocytes using MHC Class II (I-A/I-E) (M5/114.15.2) Rat mAb (redFluor™ 710 Conjugate) #29027 (solid line) compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (redFluor™ 710 Conjugate) (dashed line).
This Cell Signaling Technology antibody is conjugated to redFluor™ 710 under optimal conditions and tested in-house for direct flow cytometric analysis in mouse cells.
Note: This control antibody must be diluted to the same concentration (not dilution) as the specific antibody in analysis. See Directions for Use.
Directions for Use: Important: Dilute this control antibody to the same concentration (not dilution) as specific antibody in analysis. Higher background fluorescence may result if excessive amounts of rat IgG isotype control are used.
Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH7.2. Do not aliquot the antibody. Protect from light. Do not freeze.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc.), please refer to the dye product page for the recommended protocol. Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry.
NOTE: Count cells using a hemocytometer or alternative method.
NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to Immunostaining.
NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.
posted June 2017
revised June 2020
Protocol Id: 1504
The antibody is not directed against any known antigen. It functions as an isotype control for rat monoclonal antibodies.
Isotype control antibodies are used to estimate the nonspecific binding of target primary antibodies due to Fc receptor binding or other protein-protein interactions. An isotype control antibody should have the same immunoglobulin type and be used at the same concentration as the test antibody.