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19809
NK1.1/CD161 (PK136) Mouse mAb (redFluor™ 710 Conjugate)
Antibody Conjugates

NK1.1/CD161 (PK136) Mouse mAb (redFluor™ 710 Conjugate) #19809

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Flow cytometric analysis of live mouse splenocytes using NK1.1/CD161 (PK136) Mouse mAb (redFluor™ 710 Conjugate) and co-stained with CD3 (17A2) Rat mAb (FITC Conjugate) #86603 (right), compared to concentration-matched Mouse Isotype Control (redFluor™ 710 Conjugate) (left).

To Purchase # 19809S
Product # Size Price
19809S
100 µg $ 219

Supporting Data

REACTIVITY M
SENSITIVITY Endogenous
MW (kDa)
Isotype Mouse IgG2a, kappa

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Description

This Cell Signaling Technology antibody is conjugated to redFluor™ 710 and tested in-house for direct flow cytometric analysis in mouse cells.

Product Usage Information

For optimal flow cytometry results, we recommend 0.125 μg of antibody per test.

Application Dilution
Flow Cytometry 1:160

Storage:

Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH 7.2. This product is stable for 6 months when stored at 4ºC. Do not aliquot the antibody. Protect from light. Do not freeze.

Protocol

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Flow Cytometry, Live Cell Protocol for Directly Conjugated Antibodies

A. Solutions and Reagents

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

  1. 1X Phosphate Buffered Saline (PBS): To prepare 1 L 1X PBS: add 100 ml 10X PBS (#12528) to 900 ml water, mix.
  2. Antibody Dilution Buffer: Purchase ready-to-use Flow Cytometry Antibody Dilution Buffer (#13616), or prepare a 0.5% BSA PBS buffer by dissolving 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

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/flowdyes for a listing of cellular dyes validated for use in flow cytometry.

B. Immunostaining

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.

  1. Aliquot desired number of cells into tubes or wells. (Generally, 5x105 to 1x106 cells per assay.)
  2. Pellet cells by centrifugation and remove supernatant.
  3. Resuspend cells in 100 µl of diluted primary antibody, prepared in Antibody Dilution Buffer at a recommended dilution or as determined via titration.
  4. Incubate for 30 min to 1 hr on ice. Protect from light.
  5. Wash by centrifugation in Antibody Dilution Buffer. Discard supernatant. Repeat.
  6. Resuspend cells in 200-500 µl of Antibody Dilution Buffer and analyze on flow cytometer.

posted June 2017

revised August 2019

Protocol Id: 1504

Specificity / Sensitivity

NK1.1/CD161 (PK136) Mouse mAb (redFlour™ 710 Conjugate) recognizes endogenous levels of total NK1.1/CD161 protein. This antibody detects an epitope within the extracellular domain.

Species Reactivity:

Mouse

Source / Purification

This monoclonal antibody was purified from tissue culture supernatant via affinity chromatography. The purified antibody was conjugated under optimal conditions, with unreacted dye removed from the preparation.

Background

CD161/KLRB1 (Killer cell lectin-like receptor subfamily B member 1, also known as CLEC5B and NKR-P1A) is a type II transmembrane protein that is expressed on the majority of Natural Killer (NK) cells, NK T cells, and some T lymphocytes (1). CD161/KLRB1 is also expressed on Th17 cells, promotes their generation, and modulates their function (2). Engagement with its ligand lectin-like transcript 1 (LLT1) inhibits NK cell function, while LLT1 and CD161/KLRB1 interaction in the presence of a TCR signal enhances IFN-gamma production by T cells (3,4). There are several different CD161 isoforms in rodents and some function as activating receptors as well (5,6).

There is a family of Klrb1 genes in rodents (7). PK136 antibody recognizes a specific epitope on mouse KLRB1C and is commonly used for detection of mouse NK cells in certain mouse strains (CE, B6, NZB, C58, Ma/My, ST, SJL, FVB). However, the epitope is absent in other mouse strains (BALB/c, AKR, CBA, C3H, DBA, 129) (8).

  1. Lanier, L.L. et al. (1994) J Immunol 153, 2417-28.
  2. Bai, A. et al. (2014) J Immunol 193, 3366-77.
  3. Aldemir, H. et al. (2005) J Immunol 175, 7791-5.
  4. Rosen, D.B. et al. (2005) J Immunol 175, 7796-9.
  5. Carlyle, J.R. et al. (2006) J Immunol 176, 7511-24.
  6. Kirkham, C.L. and Carlyle, J.R. (2014) Front Immunol 5, 214.
  7. Kirkham, C.L. and Carlyle, J.R. (2014) Front Immunol 5, 214.
  8. Carlyle, J.R. et al. (2006) J Immunol 176, 7511-24.

Pathways & Proteins

Explore pathways + proteins related to this product.

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

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