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60040
KIR2DL3 (D8L3D) Rabbit mAb

KIR2DL3 (D8L3D) Rabbit mAb #60040

APPLICATIONS

REACTIVITY SENSITIVITY MW (kDa) Isotype
H Endogenous 60 Rabbit IgG
Flow Cytometry

Flow cytometric analysis of live human peripheral blood mononuclear cells using KIR2DL3 (D8L3D) Rabbit mAb compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900. Samples were co-stained with CD56 to distinguish NK cell population. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412. Total viable cells were used for analysis.

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Flow Cytometry Protocol for TIM-3 (D5D5R) XP® Rabbit mAb #45208

A. Solutions and Reagents

B. Immunostaining

NOTE: Account for isotype matched controls for monoclonal antibodies or species matched IgG for polyclonal antibodies. Count cells using a hemacytometer or alternative method.

  1. Collect cells by centrifugation and aspirate supernatant.
  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 Incubation Buffer per assay tube.
  5. Block in Incubation Buffer for 10 minutes on ice.
  6. Add the unconjugated primary antibody at the appropriate dilution to the assay tubes (see individual antibody data sheet for the dilution recommendations).
  7. Incubate for 1 hour on ice.
  8. Rinse as before in Incubation Buffer by centrifugation.
  9. Resuspend cells in fluorochrome-conjugated secondary antibody* diluted in total volume Incubation Buffer at the recommended dilution.
  10. Incubate for 30 minutes on ice.
  11. Rinse as before in Incubation Buffer by centrifugation.
  12. Resuspend cells in 0.5 ml PBS and analyze on flow cytometer.

posted August 2015

Protocol Id: 744

Application Dilutions
Flow Cytometry 1:100
Storage:

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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

Species Reactivity:

Human

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

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).

KIR2DL3, also referred to as CD158b, interacts with HLA-C alleles (HLA-Cw1, HLA-Cw3, and HLA-Cw7). Upon receptor ligand interaction, KIR2DL3 inhibits the activity of NK cells thus preventing target cell lysis (15-17).

  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.
  15. Colonna, M. et al. (1993) Proc Natl Acad Sci U S A 90, 12000-4.
  16. Winter, C.C. et al. (1998) J Immunol 161, 571-7.
  17. Moesta, A.K. et al. (2008) J Immunol 180, 3969-79.
Entrez-Gene Id
3804
Swiss-Prot Acc.
P43628
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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.

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To Purchase # 60040S

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Product # Size Price
60040S
100 µl (10 western blots) $ 255.0