Rat Focal adhesion kinase 1 (Ptk2) ELISA Kit
- SKU:
- RTEB0728
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- O35346
- ELISA Type:
- Sandwich
- Reactivity:
- Rat
Frequently bought together:
Description
| 商品名: | Rat Focal adhesion kinase 1 (Ptk2) ELISA Kit |
| 製品コード: | RTEB0728 |
| エイリアス: | Focal adhesion kinase 1, FADK 1, Protein-tyrosine kinase 2, pp125FAK, Ptk2, Fak, Fak1, 2.7.10.2, FRNK, p125FAK, pp125FAK |
| Uniprot: | O35346 |
| 反応性: | Rat |
| 範囲: | Please contact us for more information |
| 検出方法: | Sandwich |
| サイズ: | 96 Assay |
| 保管所: | Please see kit components below for exact storage details |
| ノート: | For research use only |
| UniProt Protein Function: | FAK: a tyrosine kinase of the FAK family required for cell migration and contact-dependent survival signaling. Activated by tyrosine-phosphorylation in response to either integrin clustering induced by cell adhesion or antibody cross-linking, or via G-protein coupled receptor (GPCR) occupancy by ligands such as bombesin or lysophosphatidic acid, or via LDL receptor occupancy. Downstream of integrins and Src, upstream of Ras/MAPK. Localizes to focal adhesions that form between cells growing in the presence of extracellular matrix constituents. Interacts with CAS family members and with GIT1, SORBS1 and BCAR3. Interacts with Shb. Required for full Ras transformation of fibroblasts. Increased expression in breast and other cancers, related to chromosome 8q amplification. Overexpression and activation associated with increased migration, invasion and progression of ovarian cancer, and with progression in hepatocellular carcinoma, thyroid cancer, and acute myelogenous leukemia. siRNA increases chemosensitivity of pancreatic adenocarcinoma xenografts. Inhibitor: ISI15421 (antisense). Four splice-variant isoforms have been observed.Protein type: Protein kinase, TK; Kinase, protein; EC 2.7.10.2; Protein kinase, tyrosine (non-receptor); TK group; Fak familyChromosomal Location of Human Ortholog: 8q24.3Cellular Component: extrinsic to internal side of plasma membrane; focal adhesion; cytoskeleton; lamellipodium; cytoplasm; apical plasma membrane; stress fiber; plasma membrane; microtubule organizing center; cell cortex; cytosol; nucleusMolecular Function: JUN kinase binding; protein binding; signal transducer activity; protein-tyrosine kinase activity; non-membrane spanning protein tyrosine kinase activity; SH2 domain binding; actin binding; protein kinase binding; ATP binding; receptor binding; protein kinase activityBiological Process: heart morphogenesis; axon guidance; extracellular matrix organization and biogenesis; establishment of nucleus localization; peptidyl-tyrosine phosphorylation; apoptosis; protein amino acid autophosphorylation; cell motility involved in cell locomotion; neuron migration; negative regulation of synaptogenesis; regulation of cell shape; regulation of cell adhesion mediated by integrin; transforming growth factor beta receptor signaling pathway; positive regulation of cell proliferation; ephrin receptor signaling pathway; negative regulation of axonogenesis; angiogenesis; vasculogenesis; cell structure disassembly during apoptosis; placenta development; integrin-mediated signaling pathway; epidermal growth factor receptor signaling pathway; platelet activation; regulation of osteoblast differentiation; central nervous system neuron axonogenesis; signal complex assembly; positive regulation of phosphoinositide 3-kinase activity; cytoskeleton organization and biogenesis; microtubule cytoskeleton organization and biogenesis; negative regulation of organ growth; regulation of cell proliferation; positive regulation of phosphoinositide 3-kinase cascade; positive regulation of protein kinase B signaling cascade; embryonic development; establishment of cell polarity; positive regulation of protein kinase activity; regulation of focal adhesion formation; endothelial cell migration; innate immune response; positive regulation of protein amino acid phosphorylation; negative regulation of cell-cell adhesion; blood coagulation; vascular endothelial growth factor receptor signaling pathway; positive regulation of cell migration; regulation of cytoskeleton organization and biogenesis; negative regulation of apoptosis |
| UniProt Protein Details: | |
| NCBI Summary: | This gene encodes a cytoplasmic protein tyrosine kinase which is found concentrated in the focal adhesions that form between cells growing in the presence of extracellular matrix constituents. The encoded protein is a member of the FAK subfamily of protein tyrosine kinases but lacks significant sequence similarity to kinases from other subfamilies. Activation of this gene may be an important early step in cell growth and intracellular signal transduction pathways triggered in response to certain neural peptides or to cell interactions with the extracellular matrix. Several transcript variants encoding different isoforms have been found for this gene, but the full-length natures of only three of them have been determined. [provided by RefSeq, Dec 2010] |
| UniProt Code: | O35346 |
| NCBI GenInfo Identifier: | 313851044 |
| NCBI Gene ID: | 5747 |
| NCBI Accession: | NP_001186578.1 |
| UniProt Secondary Accession: | O35346,P34152, O35346 |
| UniProt Related Accession: | O35346,Q05397 |
| Molecular Weight: | 119,233 Da |
| NCBI Full Name: | focal adhesion kinase 1 isoform c |
| NCBI Synonym Full Names: | protein tyrosine kinase 2 |
| NCBI Official Symbol: | PTK2 |
| NCBI Official Synonym Symbols: | FAK; FADK; FAK1; FRNK; PPP1R71; p125FAK; pp125FAK |
| NCBI Protein Information: | focal adhesion kinase 1; FADK 1; PTK2 protein tyrosine kinase 2; FAK-related non-kinase polypeptide; focal adhesion kinase-related nonkinase; protein phosphatase 1 regulatory subunit 71; protein phosphatase 1, regulatory subunit 71 |
| UniProt Protein Name: | Focal adhesion kinase 1 |
| UniProt Synonym Protein Names: | Focal adhesion kinase-related nonkinase; FRNK; Protein phosphatase 1 regulatory subunit 71; PPP1R71; Protein-tyrosine kinase 2; p125FAK; pp125FAK |
| Protein Family: | Focal adhesion kinase |
| UniProt Gene Name: | PTK2 |
| UniProt Entry Name: | FAK1_HUMAN |
| 成分 | 額 | 保管所 |
| 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 | - |
その他の必要な材料と設備：
- 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
*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。
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.
| ステップ | プロトコル |
| 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. |
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. |