Human Baculoviral IAP repeat-containing protein 2 / cIAP-1 ELISA Kit
- SKU:
- HUFI00959
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- Q13490
- Sensitivity:
- 46.875pg/ml
- Range:
- 78.125-5000pg/ml
- ELISA Type:
- Sandwich ELISA, Double Antibody
- Synonyms:
- BIRC2, cIAP?1, HIAP?2, MIHB, API1Hiap-2, apoptosis inhibitor 1, baculoviral IAP repeat-containing 2, baculoviral IAP repeat-containing protein 2, cIAP1, c-IAP1, hIAP2, IAP homolog B, IAP2, IAP-2, Inhibitor of apoptosis protein 2, NFR2-TRAF signalling
- Reactivity:
- Human
Description
商品名: | Human Baculoviral IAP repeat-containing protein 2 / cIAP-1 ELISA Kit |
製品コード: | HUFI00959 |
サイズ: | 96T |
エイリアス: | BIRC2, cIAP?1, HIAP?2, MIHB, API1Hiap-2, apoptosis inhibitor 1, baculoviral IAP repeat-containing 2, baculoviral IAP repeat-containing protein 2, cIAP1, c-IAP1, hIAP2, IAP homolog B, IAP2, IAP-2, Inhibitor of apoptosis protein 2, NFR2-TRAF signalling complex protein |
検出方法: | Sandwich ELISA, Double Antibody |
申し込み: | This immunoassay kit allows for the in vitro quantitative determination of Human BIRC2 concentrations in serum plasma and other biological fluids. |
感度: | 46.875pg/ml |
範囲: | 78.125-5000pg/ml |
保管所: | 4°C for 6 months |
ノート: | For Research Use Only |
回復: | Matrices listed below were spiked with certain level of Human BIRC2 and the recovery rates were calculated by comparing the measured value to the expected amount of Human BIRC2 in samples. | ||||||||||||||||
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直線性: | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human BIRC2 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. | ||||||||||||||||
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CV(%): | Intra-Assay: CV<8% Inter-Assay: CV<10% |
成分 | 量 | 保管所 |
ELISA Microplate (Dismountable) | 8×12 strips | 4°C for 6 months |
Lyophilized Standard | 2 | 4°C/-20°C |
Sample/Standard Dilution Buffer | 20ml | 4°C |
Biotin-labeled Antibody(Concentrated) | 120ul | 4°C (Protect from light) |
Antibody Dilution Buffer | 10ml | 4°C |
HRP-Streptavidin Conjugate(SABC) | 120ul | 4°C (Protect from light) |
SABC Dilution Buffer | 10ml | 4°C |
TMB Substrate | 10ml | 4°C (Protect from light) |
Stop Solution | 10ml | 4°C |
Wash Buffer(25X) | 30ml | 4°C |
Plate Sealer | 5 | - |
必要なその他の材料と設備:
- 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
Uniprot | Q13490 |
UniProt Protein Function: | BIRC2: Multi-functional protein which regulates not only caspases and apoptosis, but also modulates inflammatory signaling and immunity, mitogenic kinase signaling, and cell proliferation, as well as cell invasion and metastasis. Acts as an E3 ubiquitin- protein ligase regulating NF-kappa-B signaling and regulates both canonical and non-canonical NF-kappa-B signaling by acting in opposite directions: acts as a positive regulator of the canonical pathway and suppresses constitutive activation of non-canonical NF-kappa-B signaling. The target proteins for its E3 ubiquitin- protein ligase activity include: RIPK1, RIPK2, RIPK3, RIPK4, CASP3, CASP7, CASP8, TRAF2, DIABLO/SMAC, MAP3K14/NIK, MAP3K5/ASK1, IKBKG/NEMO and MXD1/MAD1. Can also function as an E3 ubiquitin- protein ligase of the NEDD8 conjugation pathway, targeting effector caspases for neddylation and inactivation. Acts as an important regulator of innate immune signaling via regulation of Toll-like receptors (TLRs), Nodlike receptors (NLRs) and RIG-I like receptors (RLRs), collectively referred to as pattern recognition receptors (PRRs). Protects cells from spontaneous formation of the ripoptosome, a large multi-protein complex that has the capability to kill cancer cells in a caspase-dependent and caspase-independent manner. Suppresses ripoptosome formation by ubiquitinating RIPK1 and CASP8. Can stimulate the transcriptional activity of E2F1. Plays a role in the modulation of the cell cycle. Interacts with DIABLO/SMAC and with PRSS25; these interactions inhibit apoptotic suppressor activity. Interacts with CASP9. Interacts (via BIR domains) with TRAF2. Interacts with E2F1, RIPK1, RIPK2, RIPK3, RIPK4, BIRC5/survivin and USP19. Present in many fetal and adult tissues. Mainly expressed in adult skeletal muscle, thymus, testis, ovary, and pancreas, low or absent in brain and peripheral blood leukocytes. The CARD domain inhibits the activation of E3 ubiquitin ligase activity by preventing RING domain dimerization and E2 ubiquitin donor binding and activation. The CARD domain- mediated autoinhibition of the E3 ubiquitin-protein ligase activity suppresses cell proliferation and migration. USP19 regulates the stability of BIRC2/c-IAP1 by preventing its ubiquitination. Belongs to the IAP family. |
UniProt Protein Details: | Protein type:Ubiquitin ligase; EC 6.3.2.-; Ubiquitin conjugating system; Ligase Chromosomal Location of Human Ortholog: 11q22 Cellular Component: XY body; internal side of plasma membrane; spindle microtubule; cytoplasm; nucleus; cytosol; lipid raft Molecular Function:protein binding; zinc ion binding; caspase inhibitor activity; transcription coactivator activity; ubiquitin-protein ligase activity; protein N-terminus binding; ligase activity Biological Process: proteasomal ubiquitin-dependent protein catabolic process; regulation of toll-like receptor signaling pathway; positive regulation of I-kappaB kinase/NF-kappaB cascade; protein polyubiquitination; response to cAMP; transcription, DNA-dependent; apoptosis; regulation of cell cycle; protein heterooligomerization; MyD88-independent toll-like receptor signaling pathway; toll-like receptor 3 signaling pathway; regulation of innate immune response; regulation of cell proliferation; regulation of apoptosis; response to ethanol; cell surface receptor linked signal transduction; regulation of transcription, DNA-dependent; regulation of cell differentiation; regulation of inflammatory response; response to hypoxia; toll-like receptor signaling pathway; innate immune response; toll-like receptor 4 signaling pathway; placenta development; cell structure disassembly during apoptosis; negative regulation of apoptosis |
NCBI Summary: | The protein encoded by this gene is a member of a family of proteins that inhibits apoptosis by binding to tumor necrosis factor receptor-associated factors TRAF1 and TRAF2, probably by interfering with activation of ICE-like proteases. This encoded protein inhibits apoptosis induced by serum deprivation and menadione, a potent inducer of free radicals. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jan 2012] |
UniProt Code: | Q13490 |
NCBI GenInfo Identifier: | 2497238 |
NCBI Gene ID: | 329 |
NCBI Accession: | Q13490.2 |
UniProt Secondary Accession: | Q13490,Q16516, Q4TTG0, B4E026, |
UniProt Related Accession: | Q13490 |
Molecular Weight: | 618 |
NCBI Full Name: | Baculoviral IAP repeat-containing protein 2 |
NCBI Synonym Full Names: | baculoviral IAP repeat containing 2 |
NCBI Official Symbol: | BIRC2 |
NCBI Official Synonym Symbols: | API1; MIHB; HIAP2; RNF48; cIAP1; Hiap-2; c-IAP1 |
NCBI Protein Information: | baculoviral IAP repeat-containing protein 2; IAP-2; IAP homolog B; apoptosis inhibitor 1; RING finger protein 48; inhibitor of apoptosis protein 2; baculoviral IAP repeat-containing 2; NFR2-TRAF signalling complex protein; TNFR2-TRAF-signaling complex protein 2 |
UniProt Protein Name: | Baculoviral IAP repeat-containing protein 2 |
UniProt Synonym Protein Names: | C-IAP1; IAP homolog B; Inhibitor of apoptosis protein 2; IAP-2; hIAP-2; hIAP2; RING finger protein 48; TNFR2-TRAF-signaling complex protein 2 |
Protein Family: | Baculoviral IAP repeat-containing protein |
UniProt Gene Name: | BIRC2 |
UniProt Entry Name: | BIRC2_HUMAN |
*ノート: プロトコルは、各バッチ/ロットに固有です。正しい手順については、キットに含まれているプロトコルに従ってください。
ウェルに加える前に、SABCワーキング溶液とTMB基質を37°Cで少なくとも30分間平衡化します。 サンプルと試薬を希釈するときは、完全に均一に混合する必要があります。各テストの標準曲線をプロットすることをお勧めします。
ステップ | プロトコル |
1. | Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells! |
2. | Aliquot 0.1ml standard solutions into the standard wells. |
3. | Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well. |
4. | Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells. |
5. | Seal the plate with a cover and incubate at 37 °C for 90 min. |
6. | Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2. |
7. | Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall. |
8. | Seal the plate with a cover and incubate at 37°C for 60 min. |
9. | Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash. |
10. | Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min. |
11. | Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min. |
12. | Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color. |
13. | Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately. |
14. | Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution. |
ELISAアッセイを実施する場合、可能な限り最良の結果を達成するためにサンプルを準備することが重要です。以下に、さまざまなサンプルタイプのサンプルを準備するための手順のリストを示します。
サンプルタイプ | プロトコル |
血清 | 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. |
プラズマ | 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. |
尿および脳脊髄液 | 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. |
細胞培養上清 | 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. |
細胞溶解物 | 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. |
組織ホモジネート | 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. |
組織溶解物 | 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. |
母乳 | 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. |