Human Tumor necrosis factor receptor superfamily member 14 (TNFRSF14) ELISA Kit (HUEB0557)
- SKU:
- HUEB0557
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- Q92956
- Range:
- 31.2-2000 pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- HVEM, TNFRSF14, ATAR
- Reactivity:
- Human
Description
Human Tumor necrosis factor receptor superfamily member 14 (TNFRSF14) ELISA Kit
The Human Tumor Necrosis Factor Receptor Superfamily Member 14 (TNFRSF14) ELISA Kit is specifically designed for the accurate quantification of TNFRSF14 levels in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, allowing for reliable and reproducible results that are essential for a wide range of research applications.TNFRSF14 is a critical protein belonging to the tumor necrosis factor receptor superfamily, involved in regulating immune responses and apoptosis. Its dysregulation has been associated with various autoimmune diseases, inflammatory conditions, and cancer progression.
As such, TNFRSF14 serves as a vital biomarker for studying these diseases and exploring potential therapeutic interventions.With the Human TNFRSF14 ELISA Kit, researchers can accurately measure TNFRSF14 levels in biological samples, providing valuable insights into disease mechanisms and potential treatment options. This kit is an indispensable tool for advancing research in immunology, oncology, and inflammatory disorders.
| Product Name: | Human Tumor necrosis factor receptor superfamily member 14 (TNFRSF14) ELISA Kit |
| SKU: | HUEB0557 |
| Size: | 96T |
| Target: | Human Tumor necrosis factor receptor superfamily member 14 (TNFRSF14) |
| Synonyms: | Herpes virus entry mediator A, Tumor necrosis factor receptor-like 2, Herpesvirus entry mediator A, TR2, CD270, UNQ329/PRO509, HVEA, HVEM |
| Assay Type: | Sandwich |
| Detection Method: | ELISA |
| Reactivity: | Human |
| Detection Range: | 31.2-2000pg/mL |
| Sensitivity: | 15pg/mL |
| Intra CV: | 4.2% | ||||||||||||||||||||
| Inter CV: | 7.6% | ||||||||||||||||||||
| Linearity: |
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| Recovery: |
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| Function: | (Microbial infection) Acts as a receptor for herpes simplex virus 1/HHV-1 and herpes simplex virus 1/HHV-2. |
| Uniprot: | Q92956 |
| Sample Type: | Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids |
| Specificity: | Natural and recombinant human Tumor necrosis factor receptor superfamily member 14 |
| Sub Unit: | (Microbial infection) Interacts with herpes simplex virus 1/HHV-1 and herpes simplex virus 1/HHV-2 envelope glycoprotein D (PubMed:11511370, PubMed:9696799). |
| Research Area: | Immunology |
| Subcellular Location: | Membrane Single-pass type I membrane protein |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| UniProt Protein Function: | TNFRSF14: Receptor for BTLA. Receptor for TNFSF14/LIGHT and homotrimeric TNFSF1/lymphotoxin-alpha. Involved in lymphocyte activation. Plays an important role in HSV pathogenesis because it enhanced the entry of several wild-type HSV strains of both serotypes into CHO cells, and mediated HSV entry into activated human T-cells. |
| UniProt Protein Details: | Protein type:Receptor, misc.; Membrane protein, integral; Cell cycle regulation Chromosomal Location of Human Ortholog: 1p36.32 Cellular Component: integral to plasma membrane; plasma membrane; external side of plasma membrane Molecular Function:viral receptor activity; protein binding; tumor necrosis factor receptor activity; ubiquitin protein ligase binding Biological Process: positive regulation of peptidyl-tyrosine phosphorylation; defense response to Gram-positive bacterium; entry of virus into host cell; cell surface receptor linked signal transduction; tumor necrosis factor-mediated signaling pathway; T cell costimulation; positive regulation of cytokine secretion during immune response; negative regulation of alpha-beta T cell proliferation; immune response; defense response to Gram-negative bacterium |
| NCBI Summary: | This gene encodes a member of the TNF (tumor necrosis factor) receptor superfamily. The encoded protein functions in signal transduction pathways that activate inflammatory and inhibitory T-cell immune response. It binds herpes simplex virus (HSV) viral envelope glycoprotein D (gD), mediating its entry into cells. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Jul 2014] |
| UniProt Code: | Q92956 |
| NCBI GenInfo Identifier: | 13878821 |
| NCBI Gene ID: | 8764 |
| NCBI Accession: | Q92956.3 |
| UniProt Secondary Accession: | Q92956,Q6IB95, Q8N634, Q8WXR1, Q96J31, Q9UM65, B3KW30 B9DI89, |
| UniProt Related Accession: | Q92956 |
| Molecular Weight: | 283 |
| NCBI Full Name: | Tumor necrosis factor receptor superfamily member 14 |
| NCBI Synonym Full Names: | tumor necrosis factor receptor superfamily, member 14 |
| NCBI Official Symbol: | TNFRSF14 |
| NCBI Official Synonym Symbols: | TR2; ATAR; HVEA; HVEM; CD270; LIGHTR |
| NCBI Protein Information: | tumor necrosis factor receptor superfamily member 14; CD40-like protein; herpes virus entry mediator A; tumor necrosis factor receptor-like gene2; tumor necrosis factor receptor superfamily, member 14 (herpesvirus entry mediator) |
| UniProt Protein Name: | Tumor necrosis factor receptor superfamily member 14 |
| UniProt Synonym Protein Names: | Herpes virus entry mediator A; Herpesvirus entry mediator A; HveA; Tumor necrosis factor receptor-like 2; TR2; CD_antigen: CD270 |
| Protein Family: | Tumor necrosis factor receptor superfamily |
| UniProt Gene Name: | TNFRSF14 |
| UniProt Entry Name: | TNR14_HUMAN |
| Component | Quantity (96 Assays) | Storage |
| ELISA Microplate (Dismountable) | 8×12 strips | -20°C |
| Lyophilized Standard | 2 | -20°C |
| Sample Diluent | 20ml | -20°C |
| Assay Diluent A | 10mL | -20°C |
| Assay Diluent B | 10mL | -20°C |
| Detection Reagent A | 120µL | -20°C |
| Detection Reagent B | 120µL | -20°C |
| Wash Buffer | 30mL | 4°C |
| Substrate | 10mL | 4°C |
| Stop Solution | 10mL | 4°C |
| Plate Sealer | 5 | - |
Other materials and equipment required:
- Microplate reader with 450 nm wavelength filter
- Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
- Incubator
- Deionized or distilled water
- Absorbent paper
- Buffer resevoir
*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.
| Step | |
| 1. | Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C. |
| 2. | Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform. |
| 3. | Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper. |
| 4. | Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C. |
| 5. | Repeat the wash process for five times as conducted in step 3. |
| 6. | Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction. |
| 7. | Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing. |
| 8. | Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters. |
| 9. | After experiment, store all reagents according to the specified storage temperature respectively until their expiry. |
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
| Sample Type | Protocol |
| Serum | If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. |
| Plasma | Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit. |
| Urine & Cerebrospinal Fluid | Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid. |
| Cell culture supernatant | Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately. |
| Cell lysates | Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Tissue homogenates | The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C. |
| Tissue lysates | Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Breast Milk | Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles. |
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