Description
Mouse Baculoviral IAP repeat-containing protein 5 (Birc5) ELISA Kit
The Mouse Baculoviral IAP Repeat-Containing Protein 5 (BIRC5) ELISA Kit is a highly sensitive and specific assay designed for the precise measurement of BIRC5 levels in mouse samples. This kit is ideal for the accurate detection of BIRC5 in serum, plasma, and cell culture supernatants, providing reliable and reproducible results for a variety of research applications.BIRC5, also known as survivin, is a key regulator of cell division and apoptosis, playing a crucial role in cancer development and progression.
By measuring BIRC5 levels, researchers can better understand the mechanisms underlying cancer and potentially identify new therapeutic targets for cancer treatment.With its exceptional performance and ease of use, the Mouse BIRC5 ELISA Kit is an essential tool for studying the role of BIRC5 in cancer biology and advancing our knowledge of cancer pathogenesis.
| Product Name: | Mouse Baculoviral IAP repeat-containing protein 5 (Birc5) ELISA Kit |
| SKU: | MOEB1880 |
| Size: | 96T |
| Target: | Mouse Baculoviral IAP repeat-containing protein 5 (Birc5) |
| Synonyms: | Apoptosis inhibitor 4, Apoptosis inhibitor survivin, TIAP, Api4, Iap4 |
| Assay Type: | Sandwich |
| Detection Method: | ELISA |
| Reactivity: | Mouse |
| Detection Range: | 78-5000pg/mL |
| Sensitivity: | 40.2pg/mL |
| Intra CV: | 6.6% | ||||||||||||||||||||
| Inter CV: | 11.5% | ||||||||||||||||||||
| Linearity: |
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| Recovery: |
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| Function: | Multitasking protein that has dual roles in promoting cell proliferation and preventing apoptosis. Component of a chromosome passage protein complex (CPC) which is essential for chromosome alignment and segregation during mitosis and cytokinesis. Acts as an important regulator of the localization of this complex; directs CPC movement to different locations from the inner centromere during prometaphase to midbody during cytokinesis and participates in the organization of the center spindle by associating with polymerized microtubules. Involved in the recruitment of CPC to centromeres during early mitosis via association with histone H3 phosphorylated at 'Thr-3' (H3pT3) during mitosis. The complex with RAN plays a role in mitotic spindle formation by serving as a physical scaffold to help deliver the RAN effector molecule TPX2 to microtubules. May counteract a default induction of apoptosis in G2/M phase. The acetylated form represses STAT3 transactivation of target gene promoters. May play a role in neoplasia. Inhibitor of CASP3 and CASP7. |
| Uniprot: | O70201 |
| Sample Type: | Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids |
| Specificity: | Natural and recombinant mouse Baculoviral IAP repeat-containing protein 5 |
| Sub Unit: | Monomer or homodimer. Exists as a homodimer in the apo state and as a monomer in the CPC-bound state. The monomer protects cells against apoptosis more efficiently than the dimer. Only the dimeric form is capable of enhancing tubulin stability in cells. When phosphorylated, interacts with LAMTOR5/HBXIP; the resulting complex binds pro-CASP9, as well as active CASP9, but much less efficiently. Component of the chromosomal passenger complex (CPC) composed of at least BIRC5/survivin, CDCA8/borealin, INCENP, AURKB or AURKC; in the complex forms a triple-helix bundle-based subcomplex with INCENP and CDCA8. Interacts with JTB. Interacts (via BIR domain) with histone H3 phosphorylated at 'Thr-3' (H3pT3). Interacts with EVI5. Interacts with GTP-bound RAN in both the S and M phases of the cell cycle. Interacts with USP9X. Interacts with tubulin. Interacts with BIRC2/c-IAP1. The acetylated form at Lys-129 interacts with STAT3. The monomeric form deacetylated at Lys-129 interacts with XPO1/CRM1. The monomeric form interacts with XIAP/BIRC4. Both the dimeric and monomeric form can interact with DIABLO/SMAC. Interacts with BIRC6/bruce. |
| Research Area: | Cancer |
| Subcellular Location: | Cytoplasm Nucleus Chromosome Chromosome Centromere Cytoplasm Cytoskeleton Spindle Chromosome Centromere Kinetochore Midbody Localizes on chromosome arms and inner centromeres from prophase through metaphase. Localizes to kinetochores in metaphase, distributes to the midzone microtubules in anaphase and at telophase, localizes exclusively to the midbody. Colocalizes with AURKB at mitotic chromosomes. Acetylation at Lys-129 directs its localization to the nucleus by enhancing homodimerization and thereby inhibiting XPO1/CRM1-mediated nuclear export (By similarity). |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| UniProt Protein Function: | Survivin: an apoptosis inhibitor that is expressed during the G2/M phase of the cell cycle. Associates with the microtubules of the mitotic spindle and any disruption results in the loss of apoptosis activity. May play a role in neoplasia. Inhibitor of caspase-3 and caspase-7. Two splice variant isoforms have been described. |
| UniProt Protein Details: | Protein type:Apoptosis Cellular Component: apical plasma membrane; centriole; chromosome; chromosome, pericentric region; cytoplasm; cytoplasmic microtubule; cytoskeleton; cytosol; interphase microtubule organizing center; kinetochore; microtubule; midbody; nuclear chromosome; nucleus; spindle microtubule Molecular Function:caspase inhibitor activity; chaperone binding; cofactor binding; cysteine protease inhibitor activity; enzyme binding; identical protein binding; metal ion binding; microtubule binding; protease inhibitor activity; protein binding; protein heterodimerization activity; protein homodimerization activity; Ran GTPase binding; tubulin binding; ubiquitin-protein ligase activity; zinc ion binding Biological Process: apoptosis; cell cycle; cell division; chromosome segregation; cytokinesis; embryonic development; establishment of chromosome localization; G2/M transition of mitotic cell cycle; microtubule cytoskeleton organization and biogenesis; mitosis; negative regulation of apoptosis; negative regulation of caspase activity; negative regulation of neuron apoptosis; negative regulation of peptidase activity; negative regulation of transcription, DNA-dependent; positive regulation of cell cycle; positive regulation of exit from mitosis; positive regulation of mitotic cell cycle; protein amino acid phosphorylation; protein complex localization; regulation of mitotic cell cycle; regulation of signal transduction; regulation of transcription, DNA-dependent; spindle checkpoint; transcription, DNA-dependent |
| NCBI Summary: | This gene is a member of the inhibitor of apoptosis (IAP) gene family, which encode negative regulatory proteins that prevent apoptotic cell death. IAP family members usually contain multiple baculovirus IAP repeat (BIR) domains, but this gene encodes proteins with only a single BIR domain. The encoded proteins also lack a C-terminus RING finger domain. In humans, gene expression is high during fetal development and in most tumors yet low in adult tissues. Antisense transcripts have been identified in human that regulate this gene's expression. At least three transcript variants encoding distinct isoforms have been found for this gene, although at least one of these transcript variants is a nonsense-mediated decay (NMD) candidate. [provided by RefSeq, Jul 2008] |
| UniProt Code: | O70201 |
| NCBI GenInfo Identifier: | 59859890 |
| NCBI Gene ID: | 11799 |
| NCBI Accession: | NP_001012273.1 |
| UniProt Related Accession: | O70201 |
| Molecular Weight: | 16kDa |
| NCBI Full Name: | baculoviral IAP repeat-containing protein 5 isoform 3 |
| NCBI Synonym Full Names: | baculoviral IAP repeat-containing 5 |
| NCBI Official Symbol: | Birc5 |
| NCBI Official Synonym Symbols: | Api4; TIAP; AAC-11; survivin40 |
| NCBI Protein Information: | baculoviral IAP repeat-containing protein 5 |
| UniProt Protein Name: | Baculoviral IAP repeat-containing protein 5 |
| UniProt Synonym Protein Names: | Apoptosis inhibitor 4; Apoptosis inhibitor survivin; TIAP |
| UniProt Gene Name: | Birc5 |
| UniProt Entry Name: | BIRC5_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. |
