Human Immunology ELISA Kits 5
Human Caveolin-1 (CAV1) ELISA Kit
- SKU:
- HUEB0490
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- Q03135
- Range:
- 0.156-10 ng/mL
- ELISA Type:
- Sandwich
- Synonyms:
- Cav-1, Caveolin-1, CAV1, BSCL3, CGL3, VIP21, CAV, caveolae protein, 22kD, cell growth-inhibiting protein 32,
- Reactivity:
- Human
Description
| Product Name: | Human Caveolin-1 (CAV1) ELISA Kit |
| Product Code: | HUEB0490 |
| Alias: | Caveolin-1, CAV1, CAV |
| Uniprot: | Q03135 |
| Reactivity: | Human |
| Range: | 0.156-10 ng/mL |
| Detection Method: | Sandwich |
| Size: | 96 Assay |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| UniProt Protein Function: | Caveolin-1: May act as a scaffolding protein within caveolar membranes. Interacts directly with G-protein alpha subunits and can functionally regulate their activity. Involved in the costimulatory signal essential for T-cell receptor (TCR)- mediated T-cell activation. Its binding to DPP4 induces T-cell proliferation and NF-kappa-B activation in a T-cell receptor/CD3- dependent manner. Recruits CTNNB1 to caveolar membranes and may regulate CTNNB1-mediated signaling through the Wnt pathway. Homooligomer. Interacts with GLIPR2, NOSTRIN, SNAP25 and syntaxin. Interacts with rotavirus A NSP4. Interacts (via the N- terminus) with DPP4; the interaction is direct. Interacts with CTNNB1, CDH1 and JUP. Interacts with BMX and BTK. Expressed in muscle and lung, less so in liver, brain and kidney. Belongs to the caveolin family. 2 isoforms of the human protein are produced by alternative initiation. |
| UniProt Protein Details: | Protein type:Motility/polarity/chemotaxis; Adaptor/scaffold; Nuclear receptor co-regulator Chromosomal Location of Human Ortholog: 7q31.1 Cellular Component: protein complex; focal adhesion; integral to plasma membrane; basolateral plasma membrane; endoplasmic reticulum; lipid particle; cell cortex; caveola; acrosomal membrane; cilium; lipid raft; Golgi membrane; early endosome membrane; perinuclear region of cytoplasm; apical plasma membrane; plasma membrane; intracellular; cytoplasmic vesicle; endosome Molecular Function:identical protein binding; protein binding; protease activator activity; enzyme binding; cholesterol binding; patched binding; protein complex scaffold; structural molecule activity; nitric-oxide synthase binding; protein kinase binding; receptor binding Biological Process: mammary gland involution; viral reproduction; negative regulation of epithelial cell differentiation; negative regulation of tyrosine phosphorylation of Stat5 protein; nitric oxide homeostasis; negative regulation of BMP signaling pathway; calcium ion homeostasis; protein localization; sequestering of lipid; regulation of the force of heart contraction by chemical signal; regulation of fatty acid metabolic process; negative regulation of protein binding; inactivation of MAPK activity; regulation of smooth muscle contraction; maintenance of cellular protein localization; skeletal muscle development; cytosolic calcium ion homeostasis; negative regulation of nitric-oxide synthase activity; cellular response to starvation; membrane depolarization; cholesterol homeostasis; response to estrogen stimulus; T cell costimulation; negative regulation of endothelial cell proliferation; negative regulation of protein ubiquitination; regulation of nitric-oxide synthase activity; response to calcium ion; response to progesterone stimulus; negative regulation of JAK-STAT cascade; leukocyte migration; lactation; vesicle organization and biogenesis; negative regulation of peptidyl-serine phosphorylation; negative regulation of pinocytosis; negative regulation of transcription from RNA polymerase II promoter; nitric oxide metabolic process; mammary gland development; calcium ion transport; negative regulation of cytokine and chemokine mediated signaling pathway; angiogenesis; vasculogenesis; protein homooligomerization; vasoconstriction; cholesterol transport; negative regulation of MAPKKK cascade; negative regulation of nitric oxide biosynthetic process; MAPKKK cascade; positive regulation of metalloenzyme activity; positive regulation of peptidyl-serine phosphorylation; regulation of peptidase activity; cellular calcium ion homeostasis; triacylglycerol metabolic process; regulation of blood coagulation; response to hypoxia; positive regulation of vasoconstriction; blood coagulation; vascular endothelial growth factor receptor signaling pathway Disease: Lipodystrophy, Congenital Generalized, Type 3; Partial Lipodystrophy, Congenital Cataracts, And Neurodegeneration Syndrome; Pulmonary Hypertension, Primary, 3 |
| NCBI Summary: | The scaffolding protein encoded by this gene is the main component of the caveolae plasma membranes found in most cell types. The protein links integrin subunits to the tyrosine kinase FYN, an initiating step in coupling integrins to the Ras-ERK pathway and promoting cell cycle progression. The gene is a tumor suppressor gene candidate and a negative regulator of the Ras-p42/44 mitogen-activated kinase cascade. Caveolin 1 and caveolin 2 are located next to each other on chromosome 7 and express colocalizing proteins that form a stable hetero-oligomeric complex. Mutations in this gene have been associated with Berardinelli-Seip congenital lipodystrophy. Alternatively spliced transcripts encode alpha and beta isoforms of caveolin 1.[provided by RefSeq, Mar 2010] |
| UniProt Code: | Q03135 |
| NCBI GenInfo Identifier: | 13637934 |
| NCBI Gene ID: | 857 |
| NCBI Accession: | Q03135.4 |
| UniProt Secondary Accession: | Q03135,Q9UGP1, Q9UNG1, Q9UQH6, |
| UniProt Related Accession: | Q03135 |
| Molecular Weight: | 178 |
| NCBI Full Name: | Caveolin-1 |
| NCBI Synonym Full Names: | caveolin 1, caveolae protein, 22kDa |
| NCBI Official Symbol: | CAV1 |
| NCBI Official Synonym Symbols: | CGL3; PPH3; BSCL3; LCCNS; VIP21; MSTP085 |
| NCBI Protein Information: | caveolin-1; cell growth-inhibiting protein 32 |
| UniProt Protein Name: | Caveolin-1 |
| Protein Family: | Caveolin |
| UniProt Gene Name: | CAV1 |
| UniProt Entry Name: | CAV1_HUMAN |
| 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. |
Related Products
Human CAV1 / Caveolin-1 protein ELISA Kit
