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Human NOD-like receptor family CARD domain containing 3 (NLRC3) ELISA Kit

Citations(1) Uniprot : Q7RTR2
  • Cat.No.: AE30906HU

  • Reactivity: Human (Homo sapiens)

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48T 96T
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Reagent Preparation







Results demonstration







Assay Procedure Summary







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Product Details

种属 Human (Homo sapiens)
UniProt号 Q7RTR2
缩写 NLRC3
别名 CLR16.2; FLJ00348; NOD3; CARD15-like|NOD-like receptor C3|NOD3 protein|caterpiller 16.2|nucleotide-binding oligomerization domain; leucine rich repeat and CARD domain containing 3
线性范围 0.78-50 ng/mL
灵敏度 0.39 ng/mL
样本类型 Serum, Plasma, Other biological fluids
检测方法 Sandwich
分析方法 Quantitive
反应时间 1-4.5h
样本体积 1-200 μL
检测波长 450 nm

原理

This assay employs a two-site sandwich ELISA to quantitate NLRC3 in samples. An antibody specific for NLRC3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any NLRC3 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for NLRC3 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of NLRC3 bound in the initial step. The color development is stopped and the intensity of the color is measured.
 

产品概述

CLR16.2 has a typical NBD-LRR configuration. Analysis of CLR16.2 suggests the highest expression among T lymphocytes. CLR16.2 was significantly down-regulated 6 h after anti-CD3 and anti-CD28 stimulation of primary T lymphocytes. Indeed CLR16.2 decreased NF-kappaB, NFAT, and AP-1 induction of reporter gene constructs in response to T cell activation by anti-CD3 and anti-CD28 antibodies or PMA and ionomycin. Following T cell stimulation, the presence of CLR16.2 reduced the levels of the endogenous transcripts for the IL-2 and CD25 proteins that are central in maintaining T cell activation and preventing T cell anergy. CLR16.2 serves to attenuate T cell activation via TCR and co-stimulatory molecules, and its reduction during T cell stimulation allows the ensuing cellular activation.

 

组份


Reagents

Quantity

Reagents

Quantity

Assay plate (96 Wells)

1

Instruction manual

1

Standard (lyophilized)

2

Sample Diluent

1 x 20 mL

Biotin-Conjugate (concentrate 100 x)

1 x 120 μL

Biotin-Conjugate Diluent

1 x 12 mL

Streptavidin-HRP  (concentrate 100 x)

1 x 120 μL

Streptavidin-HRP Diluent   

1 x 12 mL

Wash Buffer (concentrate 25 x)

1 x 20 mL

Substrate Solution

1 x 10 mL

Stop Solution

1 x 6 mL

Adhesive Films

4

 

特异性

This assay has high sensitivity and excellent specificity for detection of Human NLRC3. No significant cross-reactivity or interference between Human NLRC3 and analogues was observed.
 

回收率

Matrices listed below were spiked with certain level of recombinant Human NLRC3 and the recovery rates were calculated by comparing the measured value to the expected amount of Human NLRC3 in samples.
 

精密度

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%
 

线性

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human NLRC3 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
 

稳定性

The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).
 

样本收集及储存

Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2 - 8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
 

注意事项

Store at 2-8°C. Please refer to Instruction Manual.
 
  1. NLRC3: A Novel Noninvasive Biomarker for Pulmonary Hypertension Diagnosis IF: 5.058
    The nucleotide-oligomerization domain (NOD)-like receptor subfamily C3 (NLRC3) is a newly discovered and incompletely characterized member of the NLR family which negatively regulates inflammatory responses. Inflammation is considered a critical pathogenesis in pulmonary hypertension (PH).