Mouse Signal transducer and activator of transcription 2 (Stat2) ELISA Kit (MOEB1698)
- SKU:
- MOEB1698
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- Q9WVL2
- ELISA Type:
- Sandwich
- Synonyms:
- STAT2, interferon alpha induced transcriptional activator, ISGF-3
- Reactivity:
- Mouse
Description
Mouse Signal transducer and activator of transcription 2 (Stat2) ELISA Kit
The Mouse Signal Transducer and Activator of Transcription 2 (STAT2) ELISA Kit is specifically designed for the accurate detection of STAT2 levels in mouse serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring reliable and reproducible results for a variety of research applications.STAT2 is a key protein involved in the signaling pathway that regulates the immune response and interferon signaling. Dysregulation of STAT2 has been linked to autoimmune diseases, viral infections, and inflammatory conditions, making it a critical biomarker for studying these diseases and developing targeted therapies.
With its advanced technology and precise measurements, the Mouse STAT2 ELISA Kit is an essential tool for researchers investigating the role of STAT2 in immune responses and inflammatory pathways in mice. Get reliable data and valuable insights with this high-quality ELISA kit from AssayGenie.
Product Name: | Mouse Signal transducer and activator of transcription 2 (Stat2) ELISA Kit |
SKU: | MOEB1698 |
Size: | 96T |
Target: | Mouse Signal transducer and activator of transcription 2 (Stat2) |
Synonyms: | Signal transducer and activator of transcription 2, Stat2 |
Assay Type: | Sandwich |
Detection Method: | ELISA |
Reactivity: | Mouse |
Detection Range: | 0.156-10ng/mL |
Sensitivity: | 0.085ng/mL |
Intra CV: | Provided with the Kit |
Inter CV: | Provided with the Kit |
Linearity: | Provided with the Kit |
Recovery: | Provided with the Kit |
Function: | Signal transducer and activator of transcription that mediates signaling by type I IFNs (IFN-alpha and IFN-beta). Following type I IFN binding to cell surface receptors, Jak kinases (TYK2 and JAK1) are activated, leading to tyrosine phosphorylation of STAT1 and STAT2. The phosphorylated STATs dimerize, associate with IRF9/ISGF3G to form a complex termed ISGF3 transcription factor, that enters the nucleus. ISGF3 binds to the IFN stimulated response element (ISRE) to activate the transcription of interferon stimulated genes, which drive the cell in an antiviral state. Acts as a regulator of mitochondrial fission by modulating the phosphorylation of DNM1L at 'Ser-616' and 'Ser-637' which activate and inactivate the GTPase activity of DNM1L respectively. |
Uniprot: | Q9WVL2 |
Sample Type: | Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids |
Specificity: | Natural and recombinant mouse Signal transducer and activator of transcription 2 |
Sub Unit: | Heterodimer with STAT1 upon IFN-alpha/beta induced phosphorylation. The heterodimer STAT1:STAT2 forms the interferon-stimulated gene factor 3 complex (ISGF3) with IRF9; interacts with IRF9 in the cytoplasm. Interacts with CRSP2 and CRSP6. Can form a homodimer upon IFN-alpha induced phosphorylation. Interacts with IFNAR1 and IFNAR2. Interacts with ARL2BP. |
Subcellular Location: | Cytoplasm Nucleus Translocated into the nucleus upon activation by IFN-alpha/beta. |
Storage: | Please see kit components below for exact storage details |
Note: | For research use only |
UniProt Protein Function: | STAT2: transcription factor of the STAT family. Phosphorylated and activated by receptor-associated kinases downstream of many cytokines and growth-factor receptors. Binds to the IFN-stimulated response element (ISRE) and to the Gas element. This multiprotein transcription factor is termed ISGF3. Two alternatively spliced isoforms have been described. |
UniProt Protein Details: | Protein type:Transcription factor; DNA-binding Cellular Component: cytoplasm; nucleus; plasma membrane Molecular Function:DNA binding; identical protein binding; protein binding; signal transducer activity; transcription factor activity Biological Process: positive regulation of defense response to virus by host; regulation of transcription, DNA-dependent; signal transduction; transcription, DNA-dependent |
UniProt Code: | Q9WVL2 |
NCBI GenInfo Identifier: | 13633965 |
NCBI Gene ID: | 20847 |
NCBI Accession: | Q9WVL2.1 |
UniProt Secondary Accession: | Q9WVL2,Q64188, Q64189, Q64250, |
UniProt Related Accession: | Q9WVL2 |
Molecular Weight: | 105,417 Da |
NCBI Full Name: | Signal transducer and activator of transcription 2 |
NCBI Synonym Full Names: | signal transducer and activator of transcription 2 |
NCBI Official Symbol: | Stat2 |
NCBI Official Synonym Symbols: | AW496480; 1600010G07Rik |
NCBI Protein Information: | signal transducer and activator of transcription 2 |
UniProt Protein Name: | Signal transducer and activator of transcription 2 |
Protein Family: | Signal transducer and activator of transcription |
UniProt Gene Name: | Stat2 |
UniProt Entry Name: | STAT2_MOUSE |
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. |