Human Tumor necrosis factor ligand superfamily member 13 (TNFSF13) ELISA Kit (HUEB0285)
- SKU:
- HUEB0285
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- O75888
- Range:
- 0.78-50 ng/mL
- ELISA Type:
- Sandwich
- Synonyms:
- TNFSF13, APRIL, CD256
- Reactivity:
- Human
Description
Human Tumor necrosis factor ligand superfamily member 13 (TNFSF13) ELISA Kit
The Human Tumor Necrosis Factor Ligand Superfamily Member 13 (TNFSF13) ELISA Kit is specifically designed for the accurate measurement of TNFSF13 levels in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring reliable and reproducible results for a variety of research applications.TNFSF13, also known as APRIL (A Proliferation-Inducing Ligand), is a key cytokine involved in regulating immune responses and inflammation. It plays a critical role in the pathogenesis of autoimmune diseases, as well as in the growth and survival of certain types of cancer cells.
Therefore, the detection of TNFSF13 levels can provide valuable insights into disease progression and potential therapeutic targets.With its high-quality components and user-friendly protocol, the Human TNFSF13 ELISA Kit is an essential tool for researchers studying the role of TNFSF13 in various diseases and exploring new treatment strategies. Get accurate and reliable results with this innovative ELISA kit from AssayGenie.
| Product Name: | Human Tumor necrosis factor ligand superfamily member 13 (TNFSF13) ELISA Kit |
| SKU: | HUEB0285 |
| Size: | 96T |
| Target: | Human Tumor necrosis factor ligand superfamily member 13 (TNFSF13) |
| Synonyms: | A proliferation-inducing ligand, TNF- and APOL-related leukocyte expressed ligand 2, TNF-related death ligand 1, APRIL, TALL-2, TRDL-1, CD256, UNQ383/PRO715, APRIL, TALL2, ZTNF2 |
| Assay Type: | Sandwich |
| Detection Method: | ELISA |
| Reactivity: | Human |
| Detection Range: | 0.78-50ng/mL |
| Sensitivity: | 0.34ng/mL |
| Intra CV: | 5.7% | ||||||||||||||||||||
| Inter CV: | 9.4% | ||||||||||||||||||||
| Linearity: |
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| Recovery: |
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| Function: | Cytokine that binds to TNFRSF13B/TACI and to TNFRSF17/BCMA. Plays a role in the regulation of tumor cell growth. May be involved in monocyte/macrophage-mediated immunological processes. |
| Uniprot: | O75888 |
| Sample Type: | Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids |
| Specificity: | Natural and recombinant human Tumor necrosis factor ligand superfamily member 13 |
| Sub Unit: | Homotrimer. |
| Research Area: | Cancer |
| Subcellular Location: | Secreted |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| UniProt Protein Function: | TNFSF13: Cytokine that binds to TNFRSF13B/TACI and to TNFRSF17/BCMA. May be implicated in the regulation of tumor cell growth. May be involved in monocyte/macrophage-mediated immunological processes. Belongs to the tumor necrosis factor family. 4 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:Cytokine Chromosomal Location of Human Ortholog: 17p13.1 Cellular Component: cytoplasm; cytosol; external side of plasma membrane; extracellular region; extracellular space; nucleoplasm Molecular Function:cytokine activity; receptor binding; tumor necrosis factor receptor binding Biological Process: gene expression; immunoglobulin production in mucosal tissue; positive regulation of cell proliferation; positive regulation of germinal center formation; positive regulation of isotype switching to IgA isotypes; regulation of mRNA stability; signal transduction; tumor necrosis factor-mediated signaling pathway |
| NCBI Summary: | The protein encoded by this gene is a member of the tumor necrosis factor (TNF) ligand family. This protein is a ligand for TNFRSF17/BCMA, a member of the TNF receptor family. This protein and its receptor are both found to be important for B cell development. In vitro experiments suggested that this protein may be able to induce apoptosis through its interaction with other TNF receptor family proteins such as TNFRSF6/FAS and TNFRSF14/HVEM. Alternative splicing results in multiple transcript variants. Some transcripts that skip the last exon of the upstream gene (TNFSF12) and continue into the second exon of this gene have been identified; such read-through transcripts are contained in GeneID 407977, TNFSF12-TNFSF13. [provided by RefSeq, Oct 2010] |
| UniProt Code: | O75888 |
| NCBI GenInfo Identifier: | 13124605 |
| NCBI Gene ID: | 8741 |
| NCBI Accession: | O75888.1 |
| UniProt Secondary Accession: | O75888,Q541E1, Q5U0G8, Q96HV6, Q9P1M8, Q9P1M9, A8MYD5 B4DVT2, |
| UniProt Related Accession: | O75888 |
| Molecular Weight: | |
| NCBI Full Name: | Tumor necrosis factor ligand superfamily member 13 |
| NCBI Synonym Full Names: | tumor necrosis factor superfamily member 13 |
| NCBI Official Symbol: | TNFSF13 |
| NCBI Official Synonym Symbols: | APRIL; CD256; TALL2; ZTNF2; TALL-2; TNLG7B; TRDL-1; UNQ383/PRO715 |
| NCBI Protein Information: | tumor necrosis factor ligand superfamily member 13 |
| UniProt Protein Name: | Tumor necrosis factor ligand superfamily member 13 |
| UniProt Synonym Protein Names: | A proliferation-inducing ligand; APRIL; TNF- and APOL-related leukocyte expressed ligand 2; TALL-2; TNF-related death ligand 1; TRDL-1; CD_antigen: CD256 |
| UniProt Gene Name: | TNFSF13 |
| UniProt Entry Name: | TNF13_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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