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Human KIR2DL1 / CD158a HEK293 Cell Lysate (WB positive control)

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Human KIR2DL1/CD158a Transfected / Overexpression Cell Lysate Product Information
Expressed Host:Human Cells
Product Description:Human Cell lysate that Human KIR2DL1 / CD158a transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
Sequence information:A DNA sequence encoding the human KIR2DL1 (NP_055033.2) extracellular domain (Met 1-His 245) was fused with the Fc region of human IgG1 at the C-terminus.
Predicted N Terminal:His 22
Molecule Mass:The secreted recombinant human KIR2DL1/Fc is a disulfide-linked homodimer. The reduced monomer comprises 465 amino acids and has a predicted molecular mass of 51.7 kDa. The apparent molecular mass of rhKIR2DL1/Fc monomer is approximately 70 kDa in SDS-PAGE under reducing conditions due to glycosylation.
Human KIR2DL1/CD158a Transfected / Overexpression Cell Lysate Usage Guide
Preparation Method:Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer:Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Quality Control Testing:12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.
Stability:Samples are stable for up to twelve months from date of receipt.
Recommend Usage:1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
Storage Buffer:1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Storage Instruction:Store at 4℃. After re-dissolution, aliquot and store at -80℃.
Application notes:Western blot (WB): Use at an assay dependent dilution.
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
KIR2DL1/CD158a Background

Killer cell immunoglobulin-like receptor 2DL1 or KIR2DL1 is an inhibitory natural Killer cell immunoglobulin-like receptor with two extracellular immunoglobulin domains. KIR2DL1 is a member of the Killer cell immunoglobulin-like receptor family whose members are classified by the number of the extracellular immunoglobulin domains and the length of the cytoplasm domain. KIR2DL1 is a transmembrane glycoprotein expressed by natural killer cells and subsets of T cells. KIR2DL1 down-regulates the cytotoxicity of NK cells upon recognition of specific class I major histocompatibility complex (MHC) molecules on target cells. It has been reported that the KIR2DL1 bound to its class I MHC ligand, HLA-Cw4. The KIR2DL1-HLA-Cw4 interface exhibits charge and shape complementarity. Specificity is mediated by a pocket in KIR2DL1 that hosts the Lys80 residue of HLA-Cw4. Many residues conserved in HLA-C and in KIR2DL receptors make different interactions in KIR2DL1-HLA-Cw4 and in a previously reported KIR2DL2-HLA-Cw3 complex. A dimeric aggregate of KIR-HLA-C complexes was observed in one KIR2DL1-HLA-Cw4 crystal.

Human KIR2DL1/CD158a References
  • Fan QR, et al. (2001) Crystal structure of the human natural killer cell inhibitory receptor KIR2DL1-HLA-Cw4 complex. Nature Immunology. 2: 452-60.
  • Wagtmann N, et al. (1995) Molecular clones of the p58 NK cell receptor reveal immunoglobulin-related molecules with diversity in both the extra- and intracellular domains. Immunity. 2 (5): 439-49.
  • Colonna M, et al. (1995) Cloning of immunoglobulin-superfamily members associated with HLA-C and HLA-B recognition by human natural killer cells. Science. 268 (5209): 405-8.
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    Catalog: 13145-H02HL-300
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