Human TIM-3/HAVCR2 PicoKine™ Fast ELISA Kit


SKU FEK1644
Size 96wells/kit, with removable strips.
Reactivity Human
Sample Type cell culture supernates, serum and plasma (heparin, EDTA).
Sample Volume 100ul per well
Sensitivity <10pg/ml
Assay Range 31.2pg/ml-2000pg/ml

Introduction

Boster's Picokine™ Fast ELISA kits features the high sensitivity of Picokine™ ELISA kits and the significantly reduced assay run time. Now you can run the entire ELISA assay in less than 1.5 hours.

Compared to regular Picokine ELISA kits, the following sections in the datasheet are different:
1. kit components;
2. preparation before experiment;
3. assay protocol

Overview

Product Name Human TIM-3/HAVCR2 PicoKine™ Fast ELISA Kit
SKU/Catalog Number FEK1644
Storage & Handling Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)
Size 96wells/kit, with removable strips.
Description Sandwich High Sensitivity ELISA kit for Quantitative The Fast version of Picokine ELISA kits, assay takes less than 1.5 hours. Detection of Human TIM-3/HAVCR2. 96wells/kit, with removable strips.
Cite This Product Human TIM-3/HAVCR2 PicoKine™ Fast ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # FEK1644)
Sample Type cell culture supernates, serum and plasma (heparin, EDTA). Anticoagulant(s): heparin or EDTA
*The recommended anticoagulants are proven to not block the antibody binding sites on the target antigen. Please do not collect blood sample with other anticoagulants that are not specified above or contact us to check for feasibility.
Sensitivity <10pg/ml
Assay Range 31.2pg/ml-2000pg/ml
Immunogen Expression system for standard: NSO, Immunogen sequence: S22-G202
Reactivity Human
Cross Reactivity There is no detectable cross-reactivity with other relevant proteins.
Antibody Clonalities Capture Antibody | Detection Antibody:
monoclonal antibody from mouse|biotinylated polyclonal antibody from goat

Assay Details

Kit Components

Catalog number Description Quantity
FEK1644-CAP 96-well plate precoated with anti-Human HAVCR2 antibody 1
FEK1644-ST lyophilized recombinant Human HAVCR2 standard 10ng/tube
FEK1644-DA biotinylated anti-Human HAVCR2 antibody 130ul
AR1103 Avidin-Biotin-Peroxidase Complex(ABC) 130ul
AR1106-1 sample diluent buffer 30ml
AR1106-2 antibody diluent buffer 12ml
AR1106-3 ABC diluent buffer 12ml
AR1104 TMB color developing agent 10ml
AR1105 TMB stop solution 10ml
PLA-SEA Adhesive cover 4

*Why there is no wash buffer? Our Avidin-Biotin-Peroxidase Diluent contains the detergent (TWEEN) normally present in other companies' ELISA kits. This saves you the step of having to wash with the special wash buffer and achieve similar or better signal to noise ratio. The wash can use regular wash buffers (PBS, TBS etc.) commonly found in labs.

Materials Required But Not Provided

  • Microplate reader in standard size.
  • Automated plate washer.
  • Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
  • Clean tubes and Eppendorf tubes.
  • Washing buffer (neutral PBS or TBS).
  • Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g NaCl; 450μl of purified acetic acid or 700μl of concentrated hydrochloric acid to 1000ml H2
  • Preparation of 0.01 M PBS: Add 8.5g sodium chloride, 1.4g Na O and adjust pH to 7.2-7.

Typical Data Obtained from Human TIM-3/HAVCR2 PicoKine™ Fast ELISA Kit

(TMB reaction incubate at 37°C for 15-20min)

Concentration(pg/ml)02000200020002000200020002000
O.D.0.0070.0820.1570.3010.5921.0501.8622.422

Intra/Inter Assay Precision

Intra-Assay PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean(pg/ml)5122497750216884
Standard deviation2.4410.0868.392.5510.3674.25
CV(%)4.8%4.5%7%8.4%%4.8%8.4%

Reproducibility

Three samples with differing target protein concentrations were assayed using four different lots to measure the CV% lot to lot variance.

Reproducibility

To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots.

LotsLot1 (pg/ml)Lot2 (pg/ml)Lot3 (pg/ml)Lot4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 151575350522.685.1%
Sample 222422223824923310.984.7%
Sample 39771070100689898761.786.2%
*number of samples for each test n=16.

*The typical data is obtained from Boster's internal QC result and for reference only. It may differ from the lab test results of the end users. It is more important that the user's lab test results reflect the same linearity demonstrated in the typical data than achieving exactly the same O.D. values.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id Q8TDQ0
Gene Name HAVCR2
Protein Name hepatitis A virus cellular receptor 2
Tissue Specificity Expressed in T-helper type 1 (Th1) lymphocytes. Expressed on regulatory T (Treg) cells after TCR stimulation. Expressed in dendritic cells and natural killer (NK) cells. Expressed in epithelial tissues. Expression is increased on CD4+ and CD8+ T-cells in chronic hepatitis C virus (HCV) infection. In progressive HIV-1 infection, expression is up- regulated on HIV-1-specific CD8 T-cells.
Alternative Names Hepatitis A virus cellular receptor 2; HAVcr-2; T-cell immunoglobulin and mucin domain-containing protein 3; TIMD-3; T-cell immunoglobulin mucin receptor 3; TIM-3; T-cell membrane protein 3; HAVCR2; TIM3; TIMD3
Subcellular Localization Membrane ; Single-pass type I membrane protein . Cell junction

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein Function Cell surface receptor implicated in modulating innate and adaptive immune responses. Generally accepted to have an inhibiting function. Reports on stimulating functions suggest that the activity may be influenced by the cellular context and/or the respective ligand (PubMed:24825777). Regulates macrophage activation (PubMed:11823861). Inhibits T-helper type 1 lymphocyte (Th1)-mediated auto- and alloimmune responses and promotes immunological tolerance (PubMed:14556005). In CD8+ cells attenuates TCR-induced signaling, specifically by blocking NF- kappaB and NFAT promoter activities resulting in the loss of IL-2 secretion. The function may implicate its association with LCK proposed to impair phosphorylation of TCR subunits, and/or LGALS9- dependent recruitment of PTPRC to the immunological synapse (PubMed:24337741, PubMed:26492563). In contrast, shown to activate TCR-induced signaling in T-cells probably implicating ZAP70, LCP2, LCK and FYN (By similarity). Expressed on Treg cells can inhibit Th17 cell responses (PubMed:24838857). Receptor for LGALS9 (PubMed:16286920, PubMed:24337741). Binding to LGALS9 is believed to result in suppression of T-cell responses; the resulting apoptosis of antigen-specific cells may implicate HAVCR2 phosphorylation and disruption of its association with BAG6. Binding to LGALS9 is proposed to be involved in innate immune response to intracellular pathogens. Expressed on Th1 cells interacts with LGALS9 expressed on Mycobacterium tuberculosis- infected macrophages to stimulate antibactericidal activity including IL-1 beta secretion and to restrict intracellular bacterial growth (By similarity). However, the function as receptor for LGALS9 has been challenged (PubMed:23555261). Also reported to enhance CD8+ T-cell responses to an acute infection such as by Listeria monocytogenes (By similarity). Receptor for phosphatidylserine (PtSer); PtSer-binding is calcium-dependent. May recognize PtSer on apoptotic cells leading to their phagocytosis. Mediates the engulfment of apoptotic cells by dendritic cells. Expressed on T-cells, promotes conjugation but not engulfment of apoptotic cells. Expressed on dendritic cells (DCs) positively regulates innate immune response and in synergy with Toll-like receptors promotes secretion of TNF-alpha. In tumor-imfiltrating DCs suppresses nucleic acid-mediated innate immune repsonse by interaction with HMGB1 and interfering with nucleic acid-sensing and trafficking of nucleid acids to endosomes (By similarity). Expressed on natural killer (NK) cells acts as a coreceptor to enhance IFN-gamma production in response to LGALS9 (PubMed:22323453). In contrast, shown to suppress NK cell-mediated cytotoxicity (PubMed:22383801). Negatively regulates NK cell function in LPS-induced endotoxic shock (By similarity).
Research Areas Human

*You can search these to find other products in these research areas.
Background Hepatitis A virus cellular receptor 2 (HAVCR2), also known as T-cell immunoglobulin and mucin-domain containing-3 (TIM-3), is a protein that in humans is encoded by the HAVCR2 gene. The protein encoded by this gene belongs to the immunoglobulin superfamily, and TIM family of proteins. CD4-positive T helper lymphocytes can be divided into types 1 (Th1) and 2 (Th2) on the basis of their cytokine secretion patterns. Th1 cells are involved in cell-mediated immunity to intracellular pathogens and delayed-type hypersensitivity reactions, whereas, Th2 cells are involved in the control of extracellular helminthic infections and the promotion of atopic and allergic diseases. This protein is a Th1-specific cell surface protein that regulates macrophage activation, and inhibits Th1-mediated auto- and alloimmune responses, and promotes immunological tolerance.

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FEK1644
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