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

Monoclonal Antibody - NK1.1/CD161 (PK136) Mouse mAb (redFluor™ 710 Conjugate), UniProt ID P27814, Entrez ID 17059 #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

Supporting Data

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

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 Dilutions
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

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

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.