BMP6 ELISA Kit

Produktnummer ABIN624952

Produktdetails BMP6 ELISA Kit

Target: BMP6 (Bone Morphogenetic Protein 6 (BMP6))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 150-50000 pg/mL

Untere Nachweisgrenze: 150 pg/mL

Applikation: ELISA

Verwendungszweck: Human BMP-6 ELISA Kit for cell culture supernatants, plasma, and serum samples.

Proben: Plasma, Cell Culture Supernatant, Serum

Analytische Methode: Quantitative

Spezifität: This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, BMP-4, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin (OB), MCP-1, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.

Sensitivität: < 150 pg/mL

Produktmerkmale:

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

Bestandteile:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Benötigtes Material:

• Distilled or deionized water
• Precision pipettes to deliver 2 μL to 1 μL volumes
• Adjustable 1-25 μL pipettes for reagent preparation
• 100 μL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis

Anwendungsinformationen

Applikationshinweise: Recommended Dilution for serum and plasma samples2 fold

Probenmenge: 100 μL

Plattentyp: Pre-coated

Protokoll:

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 μL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Aufbereitung der Reagenzien:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
   2. Sample dilution: If your samples need to be diluted, 1x Assay Diluent (Item E) should be used for dilution of serum/plasma/culture supernatants/urine. Suggested dilution for normal serum/plasma: 2 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
   3. Assay Diluent (Item E) should be diluted 5-fold with deionized or distilled water before use.
   4. Preparation of standard: Briefly spin the vial of Item C and then add 800 µL 1x Assay Diluent (Item E, Assay Diluent should be diluted 5-fold with deionized or distilled water) into Item C vial to prepare 50,000 pg/mL standard. Dissolve the powder thoroughly by a gentle mix. Pipette 300myl 1x Assay Diluent into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Assay Diluent buffer serves as the zero standard (0 pg/mL). 200 µL 200 µL 200 µL 200 µL 200 µL Standard, Item C + 800 µL 200myl 50,000 20,000 8,000 3,200 1,280 512.0 204.8 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
   5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
   6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent and used in step 4 of Part VI Assay Procedure.
   7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 1,500-fold with 1x Assay Diluent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 8 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent to prepare a final 1,500 fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.

Testdurchführung:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
   2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
   3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
   4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
   5. Discard the solution. Repeat the wash as in step
   6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
   7. Discard the solution. Repeat the wash as in step
   8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
   9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.

Ergebnisberechnung:

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. 1x Assay Diluent Human BMP-6 concentration (pg/mL) O D =4 50 n m 0.01 0.1 1 10 100 1,000 10,000 100,000
Sensitivity: The minimum detectable dose of BMP-6 is typically less than 150 pg/mL.
Recovery: Recovery was determined by spiking various levels of human BMP-6 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 96.53 85-104 Plasma 94.69 84-103 Cell culture media 93.88 84-107
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 94 93 96 Range ( %) 85-104 84-106 87-105 1:4 Average % of Expected 95 94 94 Range ( %) 86-106 85-105 84-107
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Testpräzision: Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Handhabung: Avoid repeated freeze-thaw cycles.

Lagerung: -20 °C

Informationen zur Lagerung: The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Haltbarkeit: 6 months

Referenzen für BMP6 ELISA Kit (ABIN624952)

Zhang, Chen, Guo, Yuan, Zhang, Xu, Nemeth, Ganz, Liu: "Disordered hepcidin-ferroportin signaling promotes breast cancer growth." in: Cellular signalling, Vol. 26, Issue 11, pp. 2539-50, (2014) (PubMed).

Seamon, Wang, Cui, Keller, Dighe, Balian, Cui: "Adenoviral Delivery of the VEGF and BMP-6 Genes to Rat Mesenchymal Stem Cells Potentiates Osteogenesis." in: Bone marrow research, Vol. 2013, pp. 737580, (2013) (PubMed).

Lee, Cheng, Bilen, Lu, Satcher, Yu-Lee, Gallick, Maity, Lin: "BMP4 promotes prostate tumor growth in bone through osteogenesis." in: Cancer research, Vol. 71, Issue 15, pp. 5194-203, (2011) (PubMed).

Kim, Lee, Park, Choi, Park, Min: "The complete mitochondrial genome of the American lobster, Homarus americanus (Crustacea, Decapoda)." in: Mitochondrial DNA, Vol. 22, Issue 3, pp. 47-9, (2011) (PubMed).

Wahlström, Linder, Ansell, Kalén, Söderström, Magnusson: "Acidic preparations of lysed platelets upregulate proliferative pathways in osteoblast-like cells as demonstrated by genome-wide microarray analysis." in: Platelets, Vol. 22, Issue 6, pp. 452-60, (2011) (PubMed).

Cui, Wang, Liu, Dighe, Balian, Cui: "VEGF and BMP-6 enhance bone formation mediated by cloned mouse osteoprogenitor cells." in: Growth factors (Chur, Switzerland), Vol. 28, Issue 5, pp. 306-17, (2010) (PubMed).

Vlacic-Zischke, Hamlet, Friis, Tonetti, Ivanovski: "The influence of surface microroughness and hydrophilicity of titanium on the up-regulation of TGFβ/BMP signalling in osteoblasts." in: Biomaterials, Vol. 32, Issue 3, pp. 665-71, (2010) (PubMed).

Details zu BMP6

Target: BMP6 (Bone Morphogenetic Protein 6 (BMP6))

Andere Bezeichnung: BMP6 (BMP6 Produkte)

Synonyme: BMP6 ELISA Kit, zgc:113595 ELISA Kit, vgr ELISA Kit, vgr-1 ELISA Kit, vgr1 ELISA Kit, VGR ELISA Kit, VGR1 ELISA Kit, D13Wsu115e ELISA Kit, Vgr1 ELISA Kit, bone morphogenetic protein 6 ELISA Kit, bone morphogenetic protein 6 S homeolog ELISA Kit, BMP6 ELISA Kit, bmp6 ELISA Kit, bmp6.S ELISA Kit, Bmp6 ELISA Kit

Hintergrund: The Human BMP-6 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human BMP-6 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human BMP-6 coated on a 96-well plate. Standards and samples are pipetted into the wells and BMP-6 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human BMP-6 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of BMP-6 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.

Gen-ID: 654

UniProt: P22004

Pathways: Regulation of Hormone Metabolic Process, Regulation of Hormone Biosynthetic Process