Antibodies to NK-cells: CD335 (NKp46), CD305 (LAIR-1) and KIR – Family - FocusOn 037

Introduction: Natural killer (NK) cells are a subpopulation of lymphocytes capable of lysing transformed and virus-infected cells without apparent presensitization. Inhibitory receptors present on NK cells prevent excessive inflammatory responses and autoimmunity by binding major histocompatibility complex (MHC) expressed on target cells, resulting in the NK cells selectively killing only targets that lack MHC on their surface.

A natural cytotoxicity receptor (NCR), CD335 / NKp46, is a glycoprotein that has two extracellular Ig-like domains followed by a ~40 residue stalk region, a type I transmembrane domain, and a short cytoplasmic tail. NKp46 has been shown to represent a novel NK cell-specific molecule involved in human NK cell activation. The natural cytotoxicity receptors (NCRs) are a recently characterized family of Ig-like activation receptors that appear to be major triggering receptors in tumor cell recognition. NKp46 has been implicated in NK cell-mediated lysis of several autologous tumor cells and pathogen-infected cell lines.
CD305 / LAIR-1 is a member of the leukocyte-associated Ig like receptor family. The LAIR-1 protein shares less than 30% homology with members of the inhibitory immunoglobulin superfamily and 32% homology with members of the immunoglobulin-like transcript family. LAIR-1 is a transmembrane glycoprotein (~32kDa) expressed on the majority of human peripheral blood mononuclear leukocytes including T cells, B cells, NK cells, macrophages and dendritic cells. LAIR-1 recruits SHP-1 and SHP-2 tyrosine phosphatases upon activation, and cross-linking of the LAIR-1 molecule on NK cells results in strong inhibition of NK cell-mediated cytotoxicity.
KIR2DL1 / KIR2DL3 / KIR2DL4 / KIR2DS4 are members of the killer cell immunoglobulin-like receptor (KIR) family. The KIR family consists of up to 14 transmembrane glycoproteins of the Ig superfamily now also termed CD158a-k and CD158z. They are expressed on human NK cells and a subset of human T cells which they are involved in recognition of either MHC class I molecules or unknown ligand on target cells and inhibit cytotoxic activities. Sequence analysis revealed a structure with a 21-amino acid signal peptide, an extracellular region containing two C2-type Ig-like domains, a 19-amino acid hydrophobic transmembrane region, and a long cytoplasmic tail with 2 ITIMs.

Antibody Tools for the Detection of NK Cell Subsets

Acris Antibodies offers a broad panel of antibodies to NK cells: anti CD335 (NKp46), anti LAIR-1 and anti KIR2D antibodies can be used in flow cytometry but also in other immunological applications like ELISA or Western blotting.



Fig.1

Profile of PBMC analyzed by flow cytometry using anti NKp46 antibody SM6004.

Fig.2

Staining of human peripheral blood lymphocytes with anti human LAIR-1:FITC antibody SM2048F

Fig.3

Western blot analysis using anti KIR2DL1 antibody SM6033

Fig.1: Profile of PBMC analyzed by flow cytometry using anti NKp46 antibody SM6004.

 

Fig.2: Staining of human peripheral blood lymphocytes with anti human LAIR-1:FITC antibody SM2048F

 

Fig.3.: Western blot analysis using anti KIR2DL1 antibody SM6033: Recombinant human protein KIR2DL1, KIR2DL3, KIR2DS4 and KIR2DL4 (each 50 ng per well) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with anti-human KIR2DL1 (1:500). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.

 



Literature

  1. Warren HS. The Eighth Human Leucocyte Differentiation Antigen (HLDA8) Workshop: natural killer cell section report. Cell Immunol. 2005 Jul-Aug;236(1-2):17-20. Epub 2005 Sep 12. Erratum in: Cell Immunol. 2005 Dec;238(2):135. PMID: 16157321 [PubMed - indexed for MEDLINE]
  2. Walzer T, Bléry M, Chaix J, Fuseri N, Chasson L, Robbins SH, Jaeger S, André P, Gauthier L, Daniel L, Chemin K, Morel Y, Dalod M, Imbert J, Pierres M, Moretta A, Romagné F, Vivier E. Identification, activation, and selective in vivo ablation of mouse NK cells via NKp46. Proc Natl Acad Sci U S A. 2007 Feb 27;104(9):3384-9. Epub 2007 Feb 20
  3. Long EO, Barber DF, Burshtyn DN, Faure M, Peterson M, Rajagopalan S, Renard V, Sandusky M, Stebbins CC, Wagtmann N, Watzl C. Inhibition of natural killer cell activation signals by killer cell immunoglobulin-like receptors (CD158). Immunol Rev. 2001 Jun;181:223-33.

Available Antibody Panel to NK Cell Markers
CD335 (NKp46) antibody; CD305 (LAIR-1) antibody and LAIR-1a antibody; CD158 (KIR) antibody, CD158a (KIR2DL1), CD158b (KIR2DL3) and CD158d (KIR2DL4) antibody as well as anti - CD158i (KIR2DS4)



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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

CD158a / KIR2DL1 antibody

Western blot analysis: Recombinant Human protein KIR2DL1, KIR2DL3, KIR2DS4 and KIR2DL4 (each 50 ng per well) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with SM6033 anti-Human KIR2DL1 antibody (1/500). Proteins were visualized using a Goat anti-Mouse secondary antibody conjugated to HRP and an ECL detection system. Mouse IgG2a 2F9 Purified Hu E, IP, WB
0.1 ml / €310.00
  Acris Antibodies GmbH

CD158a / KIR2DL1 antibody

Western blot analysis: Recombinant Human protein KIR2DL1, KIR2DL3, KIR2DS4 and KIR2DL4 (each 50 ng per well) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with SM6033 anti-Human KIR2DL1 antibody (1/500). Proteins were visualized using a Goat anti-Mouse secondary antibody conjugated to HRP and an ECL detection system. Mouse IgG2a 2F9 Purified Hu E, IP, WB
50 µl / €230.00
  Acris Antibodies GmbH

CD158b / KIR2DL3 antibody

CD158b / KIR2DL3 Mouse IgG1 GL183 Purified Hu F, IP, WB
0.2 mg / €530.00
  Acris Antibodies GmbH

CD158b / KIR2DL3 antibody

CD158b / KIR2DL3 Mouse IgG2a KIR2DL3 Purified Hu F, WB
0.2 mg / €600.00
  Acris Antibodies GmbH

CD158b / KIR2DL3 antibody

CD158b / KIR2DL3 Mouse IgG2a KIR2DL3 PE Hu F
100 Tests / €590.00
  Acris Antibodies GmbH

CD158b / KIR2DL3 antibody

Profile of NK cells analyzed by Flow Cytometry Mouse IgG2a 190IIC311 Purified Hu E, F, WB
0.1 ml / €290.00
  Acris Antibodies GmbH

CD158d / KIR2DL4 (L1, L3, L4, S4) antibody

Western blot analysis: Human recombinant protein KIR2DL1, KIR2DL3 and KIR2DL4 (each 20 ng per well) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with SM6021 anti-Human KIR2DL4 antibody (1/500). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a ECL detection system.  
Lane 1 : Extracellular domain of KIR2DL1(23-223aa) 
Lane 2 : Extracellular domain of KIR2DL3(23-223aa) 
Lane 3 : Extracellular domain of KIR2DL4(44-202aa) Mouse IgG2b 2H6 Purified Hu E, WB
0.1 ml / €310.00
  Acris Antibodies GmbH

CD158d / KIR2DL4 (L1, L3, L4, S4) antibody

Western blot analysis: Human recombinant protein KIR2DL1, KIR2DL3 and KIR2DL4 (each 20 ng per well) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with SM6021 anti-Human KIR2DL4 antibody (1/500). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a ECL detection system.  
Lane 1 : Extracellular domain of KIR2DL1(23-223aa) 
Lane 2 : Extracellular domain of KIR2DL3(23-223aa) 
Lane 3 : Extracellular domain of KIR2DL4(44-202aa) Mouse IgG2b 2H6 Purified Hu E, WB
50 µl / €230.00
  Acris Antibodies GmbH

CD158i / KIR2DS4 antibody

Western blot analysis Recombinant Human KIR2DL1, KIR2DL3, KIR2DL4 and KIR2DS4 (each 100 ng) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with SM6023 anti-Human KIR2DS4 antibody (1/1000). Proteins were visualized using a Goat anti-Mouse secondary antibody conjugated to HRP and an ECL detection system. Arrow indicates recombinant Human KIR2DS4 protein. Mouse IgG2b 5F2 Purified Hu E, WB
0.1 ml / €310.00
  Acris Antibodies GmbH

CD158i / KIR2DS4 antibody

Western blot analysis Recombinant Human KIR2DL1, KIR2DL3, KIR2DL4 and KIR2DS4 (each 100 ng) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with SM6023 anti-Human KIR2DS4 antibody (1/1000). Proteins were visualized using a Goat anti-Mouse secondary antibody conjugated to HRP and an ECL detection system. Arrow indicates recombinant Human KIR2DS4 protein. Mouse IgG2b 5F2 Purified Hu E, WB
50 µl / €230.00
  Acris Antibodies GmbH

CD158 / KIR2D antibody

CD158 / KIR2D Mouse IgG1 NKVFS1 FITC Hu F
0.1 mg / €420.00
  Acris Antibodies GmbH

CD158 / KIR2D antibody

CD158 / KIR2D Mouse IgG1 NKVFS1 FITC Hu F
25 µg / €190.00
  Acris Antibodies GmbH

CD158 / KIR2D antibody

CD158 / KIR2D Mouse IgG1 NKVFS1 Low Endotoxin Hu E, F, FN, IP, WB
0.5 mg / €570.00
  Acris Antibodies GmbH

CD158 / KIR2D antibody

Staining of Human peripheral blood lymphocytes with Mouse Anti Human KIR Antibody (Cat.-No SM2036P). Mouse IgG1 NKVFS1 Aff - Purified Hu E, F, IP, WB
0.2 mg / €490.00
  Acris Antibodies GmbH

CD158 / KIR2D antibody

Staining of human peripheral blood lymphocytes with mouse anti-human KIR/KIR2D antibody Cat.-No. SM2036PS. Mouse IgG1 NKVFS1 Aff - Purified Hu E, F, IP, WB
0.1 mg / €280.00
  Acris Antibodies GmbH

CD158 / KIR2D antibody

Staining of human peripheral blood lymphocytes with mouse anti-human KIR/KIR2D antibody Cat.-No. SM2036PS. Mouse IgG1 NKVFS1 Aff - Purified Hu E, F, IP, WB
25 µg / €180.00
  Acris Antibodies GmbH

CD158 / KIR2D antibody

Staining of Human peripheral blood lymphocytes with Mouse Anti Human KIR antibody RPE conjugated (SM2036R). Mouse IgG1 NKVFS1 PE Hu F
100 Tests / €480.00
  Acris Antibodies GmbH

CD158 / KIR2D antibody

Staining of Human peripheral blood lymphocytes with Mouse Anti Human KIR antibody RPE conjugated (SM2036R). Mouse IgG1 NKVFS1 PE Hu F
25 Tests / €220.00
  Acris Antibodies GmbH

CD305 / LAIR1 antibody

Staining of Human peripheral blood lymphocytes with Mouse Anti Human CD305 antibody FITC conjugated (SM2048F). Mouse IgG1 NKTA255 FITC Hu F
0.1 mg / €380.00
  Acris Antibodies GmbH

CD305 / LAIR1 antibody

Staining of Human peripheral blood lymphocytes with Mouse Anti Human CD305 antibody FITC conjugated (SM2048F). Mouse IgG1 NKTA255 FITC Hu F
25 µg / €190.00
  Acris Antibodies GmbH

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