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76747
KIR2DL3 (D8L3D) Rabbit mAb (PE Conjugate)
Antibody Conjugates

KIR2DL3 (D8L3D) Rabbit mAb (PE Conjugate) #76747

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Flow cytometric analysis of live human peripheral blood mononuclear cells using KIR2DL3 (D8L3D) Rabbit mAb (PE Conjugate) (right), compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (PE Conjugate) #5742 (left). Samples were co-stained with NCAM1 (CD56) (MY31) Mouse mAb (APC Conjugate) #51997 to distinguish NK cell population.

To Purchase # 76747S
Product # Size Price
76747S
100 µl  (50 tests) $ 305

Supporting Data

REACTIVITY H
SENSITIVITY Endogenous
MW (kDa)
Isotype Rabbit IgG

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 phycoerythrin (PE) and tested in-house for direct flow cytometric analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated KIR2DL3 (D8L3D) Rabbit mAb #60040.

Product Usage Information

Application Dilutions
Flow Cytometry 1:50

Storage:

Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. 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

KIR2DL3 (D8L3D) Rabbit mAb (PE Conjugate) recognizes endogenous levels of total KIR2DL3 protein. This antibody weakly cross-reacts with KIR2DL2 proteins in over-expression cell lines.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala173 of human KIR2DL3 protein.

Background

Killer cell immunoglobulin-like receptors (KIRs) are type 1 transmembrane glycoproteins expressed by natural killer cells and subsets of CD4, CD8, and γδ T cells (1-5). Analogous to the diversity of their human leucocyte antigen class I (HLA Class I) ligands, the KIR genes are polymorphic and the content of the KIR gene cluster varies among haplotypes, although several "framework" genes are found in all haplotypes (6-7). The KIR proteins are characterized by the number of extracellular immunoglobulin-superfamily domains (2D or 3D) and by whether they have a long (L) or short (S) cytoplasmic domain (8-10). KIR proteins with the long cytoplasmic domain transduce inhibitory signals upon ligand binding via an immune tyrosine-based inhibitory motif (ITIM) (10), while KIR proteins with the short cytoplasmic domain lack an ITIM and instead transduce activating signals (11,12). KIR proteins play an important role in the regulation of the immune response. Combinations of KIR and HLA class I variants influence susceptibility to autoimmunity and infectious disease, as well as outcomes of haematopoietic stem cell transplantation (12-14).

  1. Young, N.T. et al. (2001) J Immunol 166, 3933-41.
  2. Battistini, L. et al. (1997) J Immunol 159, 3723-30.
  3. Björkström, N.K. et al. (2012) Blood 120, 3455-65.
  4. Remtoula, N. et al. (2008) J Immunol 180, 2767-71.
  5. Béziat, V. et al. (2017) Immunology 150, 248-264.
  6. Uhrberg, M. et al. (1997) Immunity 7, 753-63.
  7. Shilling, H.G. et al. (2002) J Immunol 168, 2307-15.
  8. Fan, Q.R. et al. (2001) Nat Immunol 2, 452-60.
  9. Boyington, J.C. et al. (2000) Nature 405, 537-43.
  10. Vivian, J.P. et al. (2011) Nature 479, 401-5.
  11. Stewart, C.A. et al. (2005) Proc Natl Acad Sci U S A 102, 13224-9.
  12. Ivarsson, M.A. et al. (2014) Front Immunol 5, 184.
  13. Kulkarni, S. et al. (2008) Semin Immunol 20, 343-52.
  14. Martin, M.P. and Carrington, M. (2013) Immunol Rev 254, 245-64.

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.
XP is a registered trademark of Cell Signaling Technology, Inc.