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40282
Mouse (MOPC-21) mAb IgG1 Isotype Control (violetFluor™ 450 Conjugate)

Mouse (MOPC-21) mAb IgG1 Isotype Control (violetFluor™ 450 Conjugate) #40282

APPLICATIONS

Concentration SENSITIVITY Isotype
200 ug/ml Endogenous Mouse IgG1
Flow Cytometry

Flow cytometric analysis of live human peripheral blood mononuclear cells using CD14 (61D3) Mouse mAb (violetFluor™ 450 Conjugate) #82944 (solid line) compared to concentration-matched Mouse (MOPC-21) mAb IgG1 Isotype Control (violetFluor™ 450 Conjugate) (dashed line). Analysis was performed on cells in the monocyte gate.

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Flow Cytometry

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (#9808) To prepare 1 L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix.
  2. Incubation Buffer: Dissolve 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

B. Immunostaining

  1. Count cells using a hemacytometer or alternative method.
  2. Aliquot 0.5-1x106 cells into each assay tube (by volume).
  3. Add 2-3 ml Incubation Buffer to each tube and rinse by centrifugation. Repeat.
  4. Resuspend cells in 100 µl of diluted primary antibody (prepared in incubation buffer at the recommended dilution).
  5. Incubate for 1 hr at room temperature.
  6. Wash by centrifugation in 2-3 ml incubation buffer.
  7. Resuspend cells in the appropriate volume of PBS and analyze on flow cytometer.

posted June 2017

Protocol Id: 1504

Note: This control antibody must be diluted to the same concentration (not dilution) as the specific antibody in analysis. See Directions on 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 mouse IgG isotype control are used. Do not use this antibody at 1:50 or 1:20 dilution like other CST violetFluor™ 450 conjugated antibodies.

Storage:

Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH7.2. This product is stable for 6 months when stored at 4ºC. Do not aliquot the antibody. Protect from light. Do not freeze.

The antibody is not directed against any known antigen. It functions as an isotype control for mouse monoclonal antibodies.

This Cell Signaling Technology antibody is conjugated to violetFluor™ 450 under optimal conditions and tested in-house for direct flow cytometric analysis in human cells.

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.

For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
violetFluor is a registered trademark of Tonbo Biosciences.

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To Purchase # 40282S

Product Number Size Price
40282S 500 µl $189.00.0
Quantity Subtotal
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