Mouse TIM-3 ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Mouse TIM-3 in Urine, Tissue Extracts, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma samples.
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Key facts
- Detection method
Colorimetric
- Sample types
Urine, Tissue Extracts, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
- Assay type
Sandwich (quantitative)
- Reactive species
Mouse
- Range
7.81 - 500 pg/mL
- Assay time
1h 30m
- Sensitivity
= 2.73 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Target data
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:18006747). Regulates macrophage activation (PubMed:11823861). Inhibits T-helper type 1 lymphocyte (Th1)-mediated auto- and alloimmune responses and promotes immunological tolerance (PubMed:14556006, PubMed:18006747). 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 (By similarity). In contrast, shown to activate TCR-induced signaling in T-cells probably implicating ZAP70, LCP2, LCK and FYN (PubMed:21807895). Expressed on Treg cells can inhibit Th17 cell responses (By similarity). Receptor for LGALS9. 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 (PubMed:22863785). 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 (PubMed:20937702). However, the function as receptor for LGALS9 has been challenged (By similarity). Also reported to enhance CD8+ T-cell responses to an acute infection such as by Listeria monocytogenes (PubMed:24567532). Receptor for phosphatidylserine (PtSer); PtSer-binding is calcium-dependent (PubMed:20083673). May recognize PtSer on apoptotic cells leading to their phagocytosis. Mediates the engulfment of apoptotic cells by dendritic cells (PubMed:19224762). Expressed on T-cells, promotes conjugation but not engulfment of apoptotic cells (PubMed:20083673). Expressed on dendritic cells (DCs) positively regulates innate immune response and in synergy with Toll-like receptors promotes secretion of TNF-alpha (PubMed:18006747). 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 (PubMed:22842346). Can enhance mast cell production of Th2 cytokines Il-4, IL-6 and IL-13 (PubMed:17620455). Expressed on natural killer (NK) cells acts as a coreceptor to enhance IFN-gamma production in response to LGALS9. In contrast, shown to suppress NK cell-mediated cytotoxicity (By similarity). Negatively regulates NK cell function in LPS-induced endotoxic shock (PubMed:25337993).
Alternative names
Hepatitis A virus cellular receptor 2 homolog, HAVcr-2, T-cell immunoglobulin and mucin domain-containing protein 3, T-cell immunoglobulin mucin receptor 3, T-cell membrane protein 3, TIMD-3, TIM-3, Havcr2, Tim3, Timd3
What's included?
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Mouse TIM-3 ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Mouse TIM-3 in Urine, Tissue Extracts, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma samples.
Key facts
- Detection method
Colorimetric
- Sample types
Urine, Tissue Extracts, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
- Assay type
Sandwich (quantitative)
- Reactive species
Mouse
- Range
7.81 - 500 pg/mL
- Assay time
1h 30m
- Assay Platform
Pre-coated microplate (12 x 8 well strips)
- Sensitivity
= 2.73 pg/mL
Precision
Intra assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Serum | n 8 | mean - | SD - | C.V. 2.3 |
Inter assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Serum | n 3 | mean - | SD - | C.V. 3 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Cell culture supernatant | Average % = 104 | Range 101 - 105 % |
Sample type Serum | Average % = 109 | Range 105 - 114 % |
Sample type EDTA Plasma | Average % = 103 | Range 102 - 106 % |
Sample type Urine | Average % = 92 | Range 91 - 94 % |
Sample type Tissue Extracts | Average % = 110 | Range 108 - 112 % |
Sample type Heparin Plasma | Average % = 103 | Range 98 - 111 % |
Sample type Citrate plasma | Average % = 108 | Range 103 - 116 % |
Storage
- Shipped at conditions
Blue Ice
- Appropriate short-term storage conditions
+4°C
- Appropriate long-term storage conditions
+4°C
- Storage information
+4°C
Notes
Mouse TIM-3 ELISA Kit (ab255721) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of TIM-3 protein in cell culture supernatant, cit plasma, edta plasma, hep plasma, serum, and urine. It uses our proprietary SimpleStep ELISA® technology. Quantitate Mouse TIM-3 with 2.73 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
TIM-3 (T-cell immunoglobulin and mucin-domain containing-3), also known as hepatitis A virus cellular receptor 2 (HAVCR2), is a type I trans-membrane protein that function as a negative regulatory immune checkpoint and can be detected in several types of immune cells. TIM-3 plays a key role in inhibiting Th1 responses and the expression of cytokines such as TNF and INF-γ. Studies have shown TIM-3 to be a promising target for cancer immunotherapy. The provided protein standard is the extracellular fragment of the full length protein.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
TIM-3 also known as T cell immunoglobulin and mucin-domain containing-3 or HAVCR2 is a protein involved in immune regulation. It possesses an approximate mass of 35 kDa. TIM-3 is expressed on various immune cells including T cells NK cells and dendritic cells especially after activation. The expression level often changes in response to inflammatory conditions suggesting its role in modulating immune responses.
- Biological function summary
TIM-3 functions as a checkpoint inhibitor impacting immune cell activity. It is not part of a larger physical complex but it modulates immune responses by interacting with its ligands such as Galectin-9 phosphatidylserine and CEACAM1. TIM-3 involvement in downregulating Th1 cell responses shows its necessary role in maintaining immune homeostasis. The protein also acts in regulating tolerance mechanisms and preventing autoimmunity.
- Pathways
TIM-3 participation is seen in the immune checkpoint and T cell exhaustion pathways. TIM-3 signaling results in T cell inhibition affecting the PD-1 pathway as well. It shares a relationship with proteins like LAG-3 and PD-1 which are key to immune inhibitory signaling. These interactions depict TIM-3's role in immune tolerance during chronic infections and malignancies.
- Associated diseases and disorders
TIM-3 association with cancer and chronic infections provides insight into therapeutic implications. In cancer TIM-3 contributes to immune evasion often co-expressed with PD-1 leading to T cell exhaustion. In autoimmune diseases TIM-3 modulation may affect disease progression by influencing immune tolerance. Understanding TIM-3's role in these contexts aids in developing targeted therapies such as anti-TIM-3 antibodies to enhance immune responses in cancer while promoting tolerance in autoimmunity.
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4 product images
Sandwich ELISA - Mouse TIM-3 ELISA Kit (ab255721)
Example of mouse TIM-3 standard curve in Sample Diluent NS.
The TIM-3 standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Mouse TIM-3 ELISA Kit (ab255721)
Interpolated concentrations of native TIM-3 in mouse serum, plasma and tissue culture supernatant samples.
The concentrations of TIM-3 were measured in duplicates, interpolated from the TIM-3 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 25%, plasma (citrate) 25%, plasma (EDTA) 50%, plasma (heparin) 50% and spleen supernatant 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TIM-3 concentration was determined to be 1045.7 pg/mL in serum, 735.6 pg/mL in plasma (citrate), 730.2 pg/mL in plasma (EDTA), 1017.5 pg/mL in plasma (heparin), and 223.3 pg/mL in spleen supernatant
Sandwich ELISA - Mouse TIM-3 ELISA Kit (ab255721)
Interpolated concentrations of native TIM-3 in mouse urine sample.
The concentrations of TIM-3 were measured in duplicates, interpolated from the TIM-3 standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 2%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TIM-3 concentration was determined to be 382.2 pg/mL in urine.
Sandwich ELISA - Mouse TIM-3 ELISA Kit (ab255721)
Interpolated concentrations of native TIM-3 in mouse spleen extract sample based on a 250 μg/mL extract load.
The concentrations of TIM-3 were measured in duplicate and interpolated from the TIM-3 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TIM-3 concentration was determined to be 43.4 pg/mL in spleen extract sample.
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