Recombinant Human CLEC-2A Protein, CF

Catalog #: 8435-CL Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
8435-CL-050

Recombinant Human CLEC-2A Protein Bioactivity

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human CLEC-2A Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE with silver staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its ability to induce IFN-gamma secretion by mouse splenocytes. The ED₅₀ for this effect is 0.4-2 μg/mL.

Source

Human embryonic kidney cell, HEK293-derived human CLEC-2A protein
Trp49-Leu174 (Gly136Asp), with an N-terminal HA tag

Accession #

Q6UVW9

N-terminal Sequence
Analysis

Tyr

Predicted Molecular Mass

16 kDa

SDS-PAGE

19-30 kDa, reducing conditions

Product Datasheets

8435-CL

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

8435-CL

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 100 μg/mL in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Bioactivity

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Recombinant Human CLEC‑2A (Catalog # 8435-CL) induces IFN-gamma secretion by mouse splenocytes. The ED₅₀ for this effect is 0.4-2 μg/mL.

SDS-PAGE

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1 μg/lane of Recombinant Human CLEC-2A was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing R bands at 22.3, 28.4 kDa and NR bands at 21.8, 29.1 kDa.

Reconstitution Calculator

Background: CLEC-2A

CLEC-2A, also called keratinocyte-associated C-type lectin (KACL), is a type 2 transmembrane glycoprotein member of the C-type lectin-like receptor (CTLR) family. CLEC-A2 is part of the subgroup of CLEC-2 proteins that also includes CLEC-2B/AICL, CLEC-2D/LLT, and CD69/CLEC-2C, all of which are encoded by the natural killer gene complex (NKC) (1, 2). CLEC-2A is composed of a 126 amino acid (aa) carboxy terminal extracellular C-type lectin-like domain (CTLD), a 21 aa transmembrane domain and a 27 aa amino terminal cytoplasmic domain. The CTLD folds into a compact structure with two alpha -helices and two antiparallel beta -sheets stabilized by three conserved intramolecular disulfide bonds (3). CLEC-2 proteins act as ligands and interact with NKRP1 receptors which also are CTLR and encoded by the NKC. The crystal structure of the complex shows CLEC-2A as a non-disulfide linked homodimer that symmetrically binds two NKp65 monomers. Interaction occurs via the membrane-distal surface of the CTLD in a head-to-head orientation (4). Five key residues for CLEC-2A binding to NKp65 are conserved or conservatively substituted among CLEC-2 proteins. Thus, the CLEC-2A-NKp65 interaction is proposed as a model for all NKRP1-CLEC-2 complexes (4). Orthologs for NKp65 and CLEC-2A are present in chimpanzee, rhesus macaque and cow but have not been described in rodents (5). Whereas CLEC-2B/AICL, CLEC-2D/LLT, and CD69/CLEC-2C are generally expressed on hematopoietic cells, CLEC-2A expression is restricted to keratinocytes. In vitro, the NKp65-CLEC-2A interaction will trigger natural killer cell cytotoxicity and the release of proinflammatory cytokines (6). Thus, CLEC-2A facilitates NKp65-mediated immunosurveillance of keratinocytes (6).

References