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Recombinant Human Kirrel1/NEPH1 His-tag Protein, CF

Catalog #: 10165-K1 Datasheet / COA / SDS
Analyzed by SEC-MALS
Catalog # Availability Size / Price Qty
10165-K1-050
Recombinant Human Kirrel1/NEPH1 His-tag Protein SEC-MALS.
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Kirrel1/NEPH1 His-tag Protein, CF Summary

Product Specifications

Purity
>95%, 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 Kirrel1/NEPH1 His-tag (Catalog # 10165-K1) is immobilized at 2 μg/mL (100 μL/well), the concentration of Recombinant Human Nephrin (Catalog # 9399-NN) that produces 50% of the optimal binding response is 1-6 μg/mL.
Source
Human embryonic kidney cell, HEK293-derived human Kirrel1/NEPH1 protein
Gln17-Leu493, with a C-terminal 6-His tag
Accession #
Q96J84
N-terminal Sequence
Analysis
Thr21
Predicted Molecular Mass
53 kDa
SDS-PAGE
65-75 kDa, under reducing conditions

Product Datasheets

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10165-K1

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.

10165-K1

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

SEC-MALS View Larger

Recombinant Human KIRREL-1/C-His Protein (Catalog # 10165-K1) has a molecular weight (MW) of 100.3 kDa as analyzed by SEC-MALS, suggesting that this protein is a homodimer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).

SDS-PAGE Recombinant Human Kirrel1/NEPH1 His-tag Protein SDS-PAGE View Larger

2 µg/lane of Recombinant Human Kirrel 1/NEPH1 His-tag (Catalog # 10165-K1) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing major bands at 65-75 kDa and 50-60 kDa, respectively.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Kirrel1/NEPH1

Kirrel1, also called NEPH1, is a 90-110 kDa type I transmembrane glycoprotein that belongs to the NEPH family of the immunoglobulin superfamily (1-4). The 757 amino acid (aa) human Kirrel1 contains a 16 aa signal sequence, a 483 aa extracellular domain (ECD) with five C2-type Ig-like domains, a 21 aa transmembrane sequence and a 237 aa cytoplasmic domain. The ECD also contains a site for FGF/FGF R interaction, and an RGD site that may allow integrin-mediated cell attachment. Five IgG-like repeats characterize the extracellular domain (5). The interaction of these five IgG-like motifs with the eight IgG-like motifs in Nephrin form a zipper-like meshwork around the glomerular capillaries in podocytes (5, 6). This interaction is what forms the structural basis of the slit diaphragm regulating macromolecule movement from the blood (5). Human Kirrel1 shares 98% aa identity with mouse and rat homologs, respectively, within the ECD. Kirrel1 expression has been mainly studied in the kidney glomerular slit diaphragm, but its expression with nephrin or other family members has also been reported in central nervous system neurons, pancreas and placenta (3, 4, 7-9). Kirrel1 forms cis hetero-oligomers with Nephrin, which brings together signaling molecules that direct actin polymerization (3, 4, 10). This interaction is essential for barrier function in the slit diaphragm, and mice deleted for Kirrel1 die perinatally due to proteinuria and failure to thrive (2, 3).

References
  1. Sellin, L. et al. (2003) FASEB J. 17:115.
  2. Donoviel, D.B. et al. (2001) Mol. Cell. Biol. 21:4829.
  3. Liu, G. et al. (2003) J. Clin. Invest. 112:209.
  4. Barletta, G-M. et al. (2003) J. Biol. Chem. 278:19266.
  5. Martin, E. and Jones, N. (2018) Front Endocrinal. 9:302.
  6. Grahammer, F. et al. (2016) JCI insight. 1: pii 86177.
  7. Gerke, P. et al. (2006) J. Comp. Neurol. 498:466.
  8. Rinta-Valkama, J. et al. (2007) Mol. Cell. Biochem. 294:117.
  9. Beall, M.H. et al. (2005) J. Soc. Gynecol. Investig. 12:298.
  10. Garg, P. et al. (2007) Mol. Cell. Biol. 27:8698.
Long Name
Kin of Irregular Chiasm-like 1
Entrez Gene IDs
55243 (Human); 170643 (Mouse); 310695 (Rat)
Alternate Names
FLJ10845; kin of IRRE like (Drosophila); KIRREL; Kirrel1; MGC129542; MGC129543; NEPH1; nephrin-like protein 1

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