Description

Nori Equine TLR2 ELISA Kit Summary

Alternative names for TLR2: Toll-like receptor 2

Alternative names for equine: horse

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: 

TLR-2 also known as CD282, is a member of the Toll-like receptor (TLR) family which plays a fundamental role in pathogen recognition and activation of innate immunity (1). TLR-2 is a membrane protein, a receptor, which recognizes foreign substances and passes on appropriate signals to the cells of the immune system. TLR-2 is expressed on microglia, Schwann cells, monocytes, macrophages, dendritic cells, polymorphonuclear leukocytes (PMNs or PMLs), B cells (B1a, MZ B, B2), and T cells, including Tregs (CD4+CD25+ regulatory T cells). In some cases it occurs in a heterodimer (combination molecule), e.g. paired with TLR-1 or TLR-6. TLR-2 is also found in the epithelia of air passages, pulmonary alveoli, renal tubules, and the Bowman’s capsules in renal corpuscles. In the skin it is found on keratinocytes and sebaceous glands. The expression of TLR2 on different cell types is regulated by different immune response modifiers. For example, LPS, GM-CSF, IL-1, and IL-10 up regulates TLR2 whereas IL-4, IFN-gamma, and TNF down regulate TLR2 expression in monocytes (2). TLR-2 recognizes many bacterial, fungal, viral, and certain endogenous substances. In general, this results in the uptake (internalization, phagocytosis) of bound molecules by endosomes/phagosomes and in cellular activation; thus such elements of innate immunity as macrophages, PMNs and dendritic cells assume functions of nonspecific immune defense, B1a and MZ B cells form the first antibodies, and specific antibody formation gets started in the process. Cytokines participating in this include tumor necrosis factor-alpha (TNF-α) and various interleukins (IL-1α, IL-1β, IL-6, IL-8, IL-12). TLR 2 has been shown to interact with TLR 1 and TOLLIP (3, 4).

References

1. Rock FL, et al (1998). Natl. Acad. Sci. U.S.A. 95 (2): 588–93.
2. Chuang TH and Ulevitch RJ.   Eur Cytokine Netw 11(3): 372-378 (2000).
3. Takeuchi, et al. (2002). Immunol. (United States) 169 (1): 10–4.
4. Zhang G and Ghosh S  (2002).  Biol. Chem. (United States) 277 (9): 7059–65.

Product Citations