Rat ROCK1 (Rhoassociated, coiled-coil containing) ELISA Kit (RTFI01306)
- SKU:
- RTFI01306
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- Q63644
- Sensitivity:
- 75pg/ml
- Range:
- 125-8000pg/ml
- ELISA Type:
- Sandwich
- Synonyms:
- p160 ROCK 1, P160ROCK, PRO0435, ROCK1
- Reactivity:
- Rat
- Research Area:
- Cell Death
Description
Rat ROCK1 (Rhoassociated, coiled-coil containing) ELISA Kit
The Rat ROCK1 (Rho-Associated, Coiled-Coil Containing Protein Kinase 1) ELISA Kit is a powerful tool for researchers studying the role of ROCK1 in various biological processes. This kit allows for the precise measurement of ROCK1 levels in rat samples, including serum, plasma, and tissue homogenates.ROCK1 is a key regulator of cell migration, invasion, and cytoskeleton dynamics, making it a crucial target for understanding diseases such as cancer, cardiovascular diseases, and neurological disorders.
By accurately quantifying ROCK1 levels, researchers can gain insights into the mechanisms underlying these conditions and potentially identify novel therapeutic targets.With its high sensitivity and specificity, the Rat ROCK1 ELISA Kit provides reliable and reproducible results, making it an essential tool for any laboratory interested in studying the role of ROCK1 in disease development and progression.
| Product Name: | Rat ROCK1 (Rhoassociated, coiled-coil containing) ELISA Kit |
| Product Code: | RTFI01306 |
| Size: | 96 Assays |
| Target: | Rat ROCK1 |
| Alias: | p160 ROCK 1, P160ROCK, PRO0435, ROCK1 |
| Reactivity: | Rat |
| Detection Method: | Sandwich ELISA, Double Antibody |
| Sensitivity: | 75pg/ml |
| Range: | 125-8000pg/ml |
| Storage: | 4°C for 6 months |
| Note: | For Research Use Only |
| Recovery: | Matrices listed below were spiked with certain level of Rat ROCK1 and the recovery rates were calculated by comparing the measured value to the expected amount of Rat ROCK1 in samples. Please contact us for more information. |
| Linearity: | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat ROCK1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Please get in contact for more information. |
| Intra-Assay: | CV <8% |
| Inter-Assay: | CV <10% |
| Uniprot: | Q63644 |
| UniProt Protein Function: | ROCK1: Protein kinase which is a key regulator of actin cytoskeleton and cell polarity. Involved in regulation of smooth muscle contraction, actin cytoskeleton organization, stress fiber and focal adhesion formation, neurite retraction, cell adhesion and motility via phosphorylation of DAPK3, GFAP, LIMK1, LIMK2, MYL9/MLC2, PFN1 and PPP1R12A. Phosphorylates FHOD1 and acts synergistically with it to promote SRC-dependent non-apoptotic plasma membrane blebbing. Phosphorylates JIP3 and regulates the recruitment of JNK to JIP3 upon UVB-induced stress. Acts as a suppressor of inflammatory cell migration by regulating PTEN phosphorylation and stability. Acts as a negative regulator of VEGF-induced angiogenic endothelial cell activation. Required for centrosome positioning and centrosome-dependent exit from mitosis. Plays a role in terminal erythroid differentiation. May regulate closure of the eyelids and ventral body wall by inducing the assembly of actomyosin bundles. Promotes keratinocyte terminal differentiation. Homodimer. Interacts with RHOB, RHOC, MYLC2B snd PTEN. Interacts with RHOA (activated by GTP), CHORDC1, DAPK3, GEM, JIP3, RHOE, PPP1R12A, PFN1, LIMK1, LIMK2 and TSG101. Interacts with FHOD1 in a Src-dependent manner. Detected in blood platelets. Activated by RHOA binding. Inhibited by Y- 27632. Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. |
| UniProt Protein Details: | Protein type:AGC group; DMPK family; EC 2.7.11.1; Kinase, protein; Protein kinase, AGC; Protein kinase, Ser/Thr (non-receptor); ROCK subfamily Chromosomal Location of Human Ortholog: 18p13 Cellular Component: cytoskeleton; lamellipodium; plasma membrane; ruffle Molecular Function:GTP-Rho binding; protein serine/threonine kinase activity Biological Process: actin cytoskeleton organization and biogenesis; bleb formation; cortical actin cytoskeleton organization and biogenesis; cytoskeleton organization and biogenesis; I-kappaB kinase/NF-kappaB cascade; leukocyte adhesion; leukocyte migration; leukocyte tethering or rolling; membrane to membrane docking; myoblast migration; negative regulation of angiogenesis; negative regulation of neuron apoptosis; negative regulation of protein binding; positive regulation of focal adhesion formation; regulation of actin cytoskeleton organization and biogenesis; regulation of actin filament-based process; regulation of keratinocyte differentiation; Rho protein signal transduction |
| NCBI Summary: | protein serine/threonine kinase; involved in intracellular signaling [RGD, Feb 2006] |
| UniProt Code: | Q63644 |
| NCBI GenInfo Identifier: | 47605939 |
| NCBI Gene ID: | 81762 |
| NCBI Accession: | Q63644.1 |
| UniProt Secondary Accession: | Q63644,Q7TP31, |
| UniProt Related Accession: | Q63644 |
| Molecular Weight: | 114,243 Da |
| NCBI Full Name: | Rho-associated protein kinase 1 |
| NCBI Synonym Full Names: | Rho-associated coiled-coil containing protein kinase 1 |
| NCBI Official Symbol: | Rock1Â Â |
| NCBI Official Synonym Symbols: | ROCK-I; P160Rock  |
| NCBI Protein Information: | rho-associated protein kinase 1 |
| UniProt Protein Name: | Rho-associated protein kinase 1 |
| UniProt Synonym Protein Names: | Liver regeneration-related protein LRRG199; Rho-associated, coiled-coil-containing protein kinase 1; Rho-associated, coiled-coil-containing protein kinase I; ROCK-I; p150 RhoA-binding kinase ROK beta; p160 ROCK-1; p160ROCK |
| Protein Family: | Rho-associated protein kinase |
| UniProt Gene Name: | Rock1Â Â |
| Step | Procedure |
| 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. |
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 clotovernight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Removeserum 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 anti-coagulant. 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 & homogenizein 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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