Human CCN5 (CCN family member 5) ELISA Kit (HUFI03415)
- SKU:
- HUFI03415
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- O76076
- Sensitivity:
- 0.094ng/ml
- Range:
- 0.156-10ng/ml
- ELISA Type:
- Sandwich
- Synonyms:
- CCN family member 5, Connective tissue growth factor-like protein, CTGF-L, Connective tissue growth factor-related protein 58, WNT1-inducible-signaling pathway protein 2, WISP-2, CCN5, CT58, CTGFL, WISP2, UNQ228, PRO261
- Reactivity:
- Human
Frequently bought together:
Description
Human CCN5 (CCN family member 5) ELISA Kit (HUFI03415)
The Human CCN5 (CCN Family Member 5) ELISA Kit is specifically designed for the quantification of CCN5 levels in human samples such as serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit guarantees accurate and reproducible results, making it an excellent choice for various research applications.CCN5, a member of the CCN family of proteins, plays a crucial role in regulating cell growth, differentiation, and migration.
Its dysregulation has been associated with various diseases, including cancer and cardiovascular disorders. The ability of this ELISA kit to accurately measure CCN5 levels makes it an invaluable tool for researchers studying these conditions and exploring potential therapeutic options.Order your Human CCN5 ELISA Kit today and advance your research with dependable and precise results.
| Product Name: | Human CCN5 (CCN family member 5) ELISA Kit |
| Product Code: | HUFI03415 |
| Size: | 96 Assays |
| Alias: | CCN family member 5 ELISA Kit, Connective tissue growth factor-like protein ELISA Kit, CTGF-L ELISA Kit, Connective tissue growth factor-related protein 58 ELISA Kit, WNT1-inducible-signaling pathway protein 2 ELISA Kit, WISP-2 ELISA Kit, CCN5 ELISA Kit, CT58 ELISA Kit, CTGFL ELISA Kit, WISP2 ELISA Kit, UNQ228 ELISA Kit, PRO261 ELISA Kit |
| Detection method: | Sandwich ELISA, Double Antibody |
| Application: | This immunoassay kit allows for the in vitro quantitative determination of Human CCN5 (CCN family member 5) concentrations in serum plasma and other biological fluids. |
| Sensitivity: | < 0.094ng/ml |
| Range: | 0.156-10ng/ml |
| Storage: | 4°C for 6 months |
| Note: | For Research Use Only |
| Recovery: | Matrices listed below were spiked with certain level of Human CCN5 (CCN family member 5) and the recovery rates were calculated by comparing the measured value to the expected amount of Human CCN5 (CCN family member 5) in samples. | ||||||||||||||||
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| Linearity: | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human CCN5 (CCN family member 5) 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% |
| Component | Quantity | Storage |
| 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 | - |
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
| UniProt Protein Function: | WISP2: May play an important role in modulating bone turnover. Promotes the adhesion of osteoblast cells and inhibits the binding of fibrinogen to integrin receptors. In addition, inhibits osteocalcin production. Belongs to the CCN family. |
| UniProt Protein Details: | Protein type:Secreted; Secreted, signal peptide Chromosomal Location of Human Ortholog: 20q13.12 Cellular Component: extracellular space; proteinaceous extracellular matrix; plasma membrane Molecular Function:heparin binding; integrin binding; insulin-like growth factor binding Biological Process: cell-cell signaling; regulation of cell growth; signal transduction; cell adhesion |
| NCBI Summary: | This gene encodes a member of the WNT1 inducible signaling pathway (WISP) protein subfamily, which belongs to the connective tissue growth factor (CTGF) family. WNT1 is a member of a family of cysteine-rich, glycosylated signaling proteins that mediate diverse developmental processes. The CTGF family members are characterized by four conserved cysteine-rich domains: insulin-like growth factor-binding domain, von Willebrand factor type C module, thrombospondin domain and C-terminal cystine knot-like (CT) domain. The encoded protein lacks the CT domain which is implicated in dimerization and heparin binding. It is 72% identical to the mouse protein at the amino acid level. This gene may be downstream in the WNT1 signaling pathway that is relevant to malignant transformation. Its expression in colon tumors is reduced while the other two WISP members are overexpressed in colon tumors. It is expressed at high levels in bone tissue, and may play an important role in modulating bone turnover. [provided by RefSeq, Jul 2008] |
| UniProt Code: | O76076 |
| NCBI GenInfo Identifier: | 24211561 |
| NCBI Gene ID: | 8839 |
| NCBI Accession: | O76076.1 |
| UniProt Secondary Accession: | O76076,Q6PEG3, B2R9N4, E1P612, |
| UniProt Related Accession: | O76076 |
| Molecular Weight: | Observed: 27kDaCalculated: 22kDa/26kDa |
| NCBI Full Name: | WNT1-inducible-signaling pathway protein 2 |
| NCBI Synonym Full Names: | WNT1 inducible signaling pathway protein 2 |
| NCBI Official Symbol: | WISP2 |
| NCBI Official Synonym Symbols: | CCN5; CT58; CTGF-L |
| NCBI Protein Information: | WNT1-inducible-signaling pathway protein 2; WISP-2; CCN family member 5; wnt-1 signaling pathway protein 2; connective tissue growth factor-like protein; connective tissue growth factor-related protein 58 |
| UniProt Protein Name: | WNT1-inducible-signaling pathway protein 2 |
| UniProt Synonym Protein Names: | CCN family member 5; Connective tissue growth factor-like protein; CTGF-L; Connective tissue growth factor-related protein 58 |
| Protein Family: | WNT1-inducible-signaling pathway protein |
| UniProt Gene Name: | WISP2 |
| UniProt Entry Name: | WISP2_HUMAN |
*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37 °C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.
| Step | Protocol |
| 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 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. |
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