Recombinant Human MICL/CLEC12A Fc Chimera Protein, CF

Catalog #: 10835-ML Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
10835-ML-050

Recombinant Human MICL/CLEC12A Fc Chimera Protein Binding Activity.

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

Recombinant Human MICL/CLEC12A Fc Chimera Protein, CF Summary

Product Specifications

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

Activity

Measured by its binding ability in a functional ELISA. When Recombinant Human Integrin alpha X beta 2 Protein (Catalog # 5755-AX) is immobilized at 5.00 μg/mL, 100 μL/well, the concentration of Recombinant Human MICL/CLEC12A Fc Chimera (Catalog# 10835-ML) that produces 50% of the optimal binding response is approximately 1.20-12.0 μg/mL.

Source

Chinese Hamster Ovary cell line, CHO-derived human MICL/CLEC12A protein

MD	Human IgG₁ (Pro100-Lys330)	IEGR	Human MICL/CLEC12A (Thr67-Ala265) Accession # AAI26292.1
N-terminus			C-terminus

Accession #

AAI26292.1

N-terminal Sequence
Analysis

Met

Structure / Form

Disulfide-linked homodimer

Predicted Molecular Mass

50 kDa

SDS-PAGE

70-85 kDa, under reducing conditions

Product Datasheets

10835-ML

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.

10835-ML

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 500 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

Binding Activity

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When Recombinant Human Integrin alpha X beta 2 Protein (5755-AX) immobilized at 5.00 μg/mL, 100 μL/well, the concentration of Recombinant Human MICL/CLEC12A Fc Chimera (Catalog# 10835-ML) that produces 50% of the optimal binding response is approximately 1.20-12.0 μg/mL.

SDS-PAGE

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2 μg/lane of Recombinant Human MICL/CLEC12A Fc Chimera (Catalog # 10835-ML) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 70-85 kDa and 140-170 kDa, respectively.

Reconstitution Calculator

Background: MICL/CLEC12A

C-type lectin domain family 12 member A (CLEC12A), also known as C-type lectin-like molecule 1 (CLL1), dendritic cell-associated lectin 2 (DCAL-2), myeloid inhibitory C-type lectin-like receptor (MICL), killer cell lectin like receptor-1 (KLRL1) and CD371, is a member of the C-type lectin receptor superfamily. Of the 17 groups in the superfamily, CLEC12A belongs to group V, which lacks the domain for calcium binding typically found in classical carbohydrate-binding CLECs (1). Mature CLEC12A consists of an extracellular domain (ECD) with a single C-type lectin-like domain, a type II transmembrane domain and a cytoplasmic tail containing an immunoreceptor tyrosine-based inhibition motif (ITIM) (3). Within the ECD, human CLEC12A shares 50% and 47% amino acid sequence identity with mouse and rat CLEC12A, respectively. Human CLEC12A differs from mouse CLEC12A in that it is heavily glycosylated and is found as a monomer rather than a dimer (2). CLEC12A is predominantly expressed in innate immune cells and plays a role in immunotherapy (1-6). CLEC12A preferentially associates with the protein tyrosine phosphatases SHP-1 and SHP-2 but not SHIP. Mechanistic studies with chimeric proteins have indicated that, similar to other ITIM-containing receptors, the cytoplasmic tail of CLEC12A can inhibit cellular activation upon ligand binding stimulation. The physiological functions of CLEC12A are poorly understood and the human ligand for CLEC12A is unknown. (7) Most CTL receptors require Ca2+ ions for binding (8). C-type lectins are the most diverse and prevalent lectin family in immunity. Particular interest has recently been attracted by the C-type lectin-like receptors on NK cells, which appear to regulate the activation/inhibitory balance of these cells, controlling cytotoxicity and cytokine production (9). CLEC12A receptor has emerged as a leukemia-associated and cancer stem cell marker in myeloid malignancies (10). CLEC12A is a myeloid lineage antigen that is highly expressed by AML cells and LSCs, but not expressed by normal hematopoietic stem cells (HSCs). The CLEC12A TriKE induced robust NK-cell specific proliferation, enhanced NK-cell activation, and killing of both AML cell lines and primary patient-derived AML blasts in vitro while sparing healthy HSCs. Additionally, the CLEC12A TriKE was able to reduce tumor burden in preclinical mouse models. These findings highlight the clinical potential of the CLEC12A TriKE for the effective treatment of AML (11).

References