Description

Nori Human TLR1 ELISA Kit Summary

Alternative names for TLR1: Toll-like receptor 1 (TLR1), CD281

Q15399

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background:

TLR1 is a member of the Toll-like receptor family (TLR) of pattern recognition receptors of the innate immune system (1,2). TLR1 recognizes pathogen-associated molecular pattern with a specificity for gram-positive bacteria. TLR1 has also been designated as CD281 (cluster of differentiation 281).

TLRs TLRs contain a large number of leucinerich repeats (LRRs) and a cytoplasmic tail with one Toll/IL1 receptor (TIR) domain.They are type I transmembrane proteins that serve as pattern recognition receptors for microbial pathogens,  and highly conserved from Drosophila to humans and share structural and functional similarities. They recognize pathogen-associated molecular patterns (PAMPs) that are expressed on infectious agents, and mediate the production of cytokines necessary for the development of effective immunity. The various TLRs exhibit different patterns of expression. This gene is ubiquitously expressed, and at higher levels than other TLR genes. Different length transcripts presumably resulting from use of alternative polyadenylation site, and/or from alternative splicing, have been noted for this gene (3).

TLR1 recognises peptidoglycan and (triacyl) lipoproteins in concert with TLR2 (as a heterodimer) (4, 5). It is found on the surface of macrophages and neutrophils. TLR 1 has been shown to interact with TLR 2 (6).

References

1. Rock FL, et al. Proc. Natl. Acad. Sci. U.S.A. 95 (2): 588–93.
2. Lien E, et al. Crit. Care Med. 30 (1 Suppl): S1–11.
3. “SRF serum response factor”. Entrez Gene. National Center for Biotechnology Information, National Institutes of Health.
4. Farhat K, et al. J. Leukoc. Biol. 83 (3): 692–701.
5. Jin MS, et al. Cell 130 (6): 1071–82.
6. Osamu T, et al. J. Immunol. (United States) 169 (1): 10–4.

Product Citations