COL2 ELISA Kit

Produktnummer ABIN6954923

Produktdetails COL2 ELISA Kit

Target: COL2 (Collagen, Type II (COL2))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 93.75 pg/mL - 6000 pg/mL

Untere Nachweisgrenze: 93.75 pg/mL

Applikation: ELISA

Verwendungszweck: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of COL2 in human serum, plasma, tissue homogenates.

Proben: Plasma, Serum, Tissue Homogenate

Analytische Methode: Quantitative

Spezifität: This assay has high sensitivity and excellent specificity for detection of Collagen Type II (COL2)

Sensitivität: 34.82 pg/mL

Bestandteile:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Anwendungsinformationen

Kommentare:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Probenmenge: 100 μL

Testdauer: 3 h

Plattentyp: Pre-coated

Protokoll:

1. Prepare all reagents, samples and standards,
2. Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
4. Aspirate and wash 3 times,
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
6. Aspirate and wash 5 times,
7. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
8. Add 50μL Stop Solution. Read at 450nm immediately.

Aufbereitung der Reagenzien:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 12,000pg/mL. Firstly dilute the stock solution to 6,000pg/mL and the diluted standard serves as the highest standard (6,000pg/mL). Then prepare 7 tubes containing 0.5 mL Standard Diluent and use the diluted standard to produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 6,000pg/mL, 3,000pg/mL, 1,500pg/mL, 750pg/mL, 375pg/mL, 187.5pg/mL, 93.75pg/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0pg/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Aufbereitung der Proben:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Testpräzision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Vorsichtsmaßnahmen: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Lagerung: 4 °C/-20 °C

Informationen zur Lagerung:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

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Haltbarkeit: 6 months

Referenzen für COL2 ELISA Kit (ABIN6954923)

Samuel, Ahmad, Ramasamy, Karunanithi, Naveen, Kamarul et al.: "Platelet-rich concentrate in serum-free medium enhances cartilage-specific extracellular matrix synthesis and reduces chondrocyte hypertrophy of human mesenchymal stromal cells encapsulated in ..." in: Platelets, Vol. 30, Issue 1, pp. 66-74, (2019) (PubMed).

Mern, Tschugg, Hartmann, Thomé et al.: "Self-complementary adeno-associated virus serotype 6 mediated knockdown of ADAMTS4 induces long-term and effective enhancement of aggrecan in degenerative human nucleus pulposus cells: A new ..." in: PLoS ONE, Vol. 12, Issue 2, pp. e0172181, (2017) (PubMed).

Gualandi, Torricelli, Panzavolta, Pagani, Focarete, Bigi: "An innovative co-axial system to electrospin in situ crosslinked gelatin nanofibers." in: Biomedical materials (Bristol, England), Vol. 11, Issue 2, pp. 025007, (2016) (PubMed).

Krouwels, Popov-Celeketic, Plomp, Dhert, Öner, Bank, Creemers: "No effects of Hyperosmolar Culture medium on Tissue Regeneration by Human Degenerated Nucleus Pulposus Cells despite Upregulation Extracellular Matrix Genes." in: Spine, (2015) (PubMed).

Amadori, Torricelli, Panzavolta, Parrilli, Fini, Bigi: "Highly Porous Gelatin Reinforced 3D Scaffolds for Articular Cartilage Regeneration." in: Macromolecular bioscience, Vol. 15, Issue 7, pp. 941-52, (2015) (PubMed).

Xu, Ke, Wang, Lin: "The role of MCP-1-CCR2 ligand-receptor axis in chondrocyte degradation and disease progress in knee osteoarthritis." in: Biological research, Vol. 48, pp. 64, (2015) (PubMed).

Park, Park, Chung, O, Waner: "Comparative Analysis of the Extracellular Matrix Composition in Proliferating and Involuted Infantile Hemangiomas." in: Archives of plastic surgery, Vol. 42, Issue 5, pp. 544-51, (2015) (PubMed).

Yang, Hao, Hu: "MicroRNA expression profiles in human adipose-derived stem cells during chondrogenic differentiation." in: International journal of molecular medicine, Vol. 35, Issue 3, pp. 579-86, (2015) (PubMed).

Amable, Teixeira, Carias, Granjeiro, Borojevic: "Protein synthesis and secretion in human mesenchymal cells derived from bone marrow, adipose tissue and Wharton's jelly." in: Stem cell research & therapy, Vol. 5, Issue 2, pp. 53, (2014) (PubMed).

Torricelli, Gioffrè, Fiorani, Panzavolta, Gualandi, Fini, Focarete, Bigi: "Co-electrospun gelatin-poly(L-lactic acid) scaffolds: modulation of mechanical properties and chondrocyte response as a function of composition." in: Materials science & engineering. C, Materials for biological applications, Vol. 36, pp. 130-8, (2014) (PubMed).

Amable, Teixeira, Carias, Granjeiro, Borojevic: "Mesenchymal stromal cell proliferation, gene expression and protein production in human platelet-rich plasma-supplemented media." in: PLoS ONE, Vol. 9, Issue 8, pp. e104662, (2014) (PubMed).

Li, Wang, Zhang, Zhao, Huang, Wu, Li, Li, Liu, Cao, Dai, Fang, Shang, Cao, Zhao, Chen: "Elevated PLGF contributes to small-cell lung cancer brain metastasis." in: Oncogene, Vol. 32, Issue 24, pp. 2952-62, (2013) (PubMed).

Mern, Fontana, Beierfuß, Thomé, Hegewald et al.: "A combinatorial relative mass value evaluation of endogenous bioactive proteins in three-dimensional cultured nucleus pulposus cells of herniated intervertebral discs: identification of potential ..." in: PLoS ONE, Vol. 8, Issue 11, pp. e81467, (2013) (PubMed).

Li, Liang, Tao, Zhou, Li, Chen, Chen: "Acidic pH conditions mimicking degenerative intervertebral discs impair the survival and biological behavior of human adipose-derived mesenchymal stem cells." in: Experimental biology and medicine (Maywood, N.J.), Vol. 237, Issue 7, pp. 845-52, (2012) (PubMed).

Details zu COL2

Target: COL2 (Collagen, Type II (COL2))

Andere Bezeichnung: Collagen Type II (COL2) (COL2 Produkte)