Humain BMP-2 ELISA Pair Set

Détails pour le produit réf. ABIN2010205
Antigène
  • BDA2
  • BMP2A
  • AI467020
  • Bmp2a
  • BMP-2
  • xBMP-2
  • xbmp2
  • BMP2
  • bmp2a
  • bmp2
  • wu:fc59d09
  • bone morphogenetic protein 2
  • bone morphogenetic protein 2 L homeolog
  • Bone morphogenetic protein 2
  • bone morphogenetic protein 2a
  • BMP2
  • Bmp2
  • bmp2.L
  • bmp2
  • bmp2a
Reactivité
Humain
Méthode de détection
Colorimetric
Hôte
Souris
Conjugué
HRP
Type de méthode
Sandwich ELISA
Application
ELISA
Options
Fonction The Human BMP-2 ELISA Pair Set is for the quantitative determination of Human BMP-2.
This ELISA Pair Set contains the basic components required for the development of sandwich ELISAs.
Méthode de détection Colorimetric
Sensibilité 0.03125 pg/mL
Attributs du produit Capture Antibody: 1.0 mg/mL of mouse anti-BMP2 monoclonal antibody, Dilute to a working concentration of 2.0 μg/mL in CBS before coating.

Detection Antibody: 0.5 mg/mL mouse anti-BMP2 monoclonal antibody conjugated to horseradish-peroxidase (HRP). Dilute to working concentration of 0.25 μg/mL in detection antibody dilution buffer before use.

Standard: Each vial contains 30 ng of recombinant BMP2. Reconstitute standard powder with 1 mL detection antibody dilution buffer. After reconstitution, store at -20 °C to -80 °C in a manual defrost freezer. A seven-point standard curve usi ng 2-fold serial dilutions in sample dilution buffer, and a high standard of 2000 pg/mL is recommended.

Sensitivity: The minimum detectable dose of Human BMP-2 was determined to be approximately 0.03125 pg/mL. This is defined as at least three times standard deviations above the mean optical density of 10 replicates of the zero standard.
Ingrédients
  • Capture Ab
  • Detection Ab
  • Standard
Matériel non inclus CBS - 0.05 M
Na2CO3 - 0.05 M
NaHCO3, pH 9.6, 0.2 μm filtered
TBS - 25 mM
Tris, adjust pH to 7.4 by HCl
Wash Buffer - 0.05 % Tween20 in TBS, pH 7.2 - 7.4
Blocking Buffer - 5 % BSA in Wash Buffer
Sample dilution buffer - 0.5 % BSA in wash buffer, pH 7.2 - 7.4, 0.2 μm filtered
Detection antibody dilution buffer - 0.5 % BSA in wash buffer, pH 7.2 - 7.4, 0.2 μm filtered
Substrate Solution: To achieve best assay results, fresh substrate solution is recommended Substrate stock solution - 10 mg/mL TMB (Tetramethylbenzidine ) in DMSO Substrate dilution buffer - 0.05 M Na2HPO4 and 0.025 M citric acid , adjust pH to 5.5
Substrate working solution - For each plate dilute 250 μL substrate stock solution in 25 mL substrate dilutionbuffer and then add 80 μL 0.75 % H2O2 , mix it well
Stop Solution - 2 N H2SO4
Antigène
Autre désignation BMP2
Sujet Bone morphogenetic protein 2 (BMP2) is a member of the BMP subgroup belonging to the transforming growth factor(TGF) -beta superfamily. The mature BMP2 acts as disulfide-linked homodimers and heterodimers with BMP-7 afterproteolytic removal of propeptide. BMP2 is capable of inducing the formation of cartilage and bone but are nowregarded as multifunctional cytokines. It binds heterodimeric receptor complexes composed of a type I receptor and atype I I receptor, and transduces the related signals via the down-stream molecules, such as the PI3 kinase, Smads, STATs, etc. BMP2 has been demonstrated to potently induce osteoblast differentiation in a variety of cell types, andinduce apoptosis in human myeloma cell lines as a novel function. In addition, recent studies have revealed thatosteoblast-derived BMP2 enhances the migration of prostate cancer cells. rhBMP2 can be utilized in various therapeuticinterventions such as bony defects, delayed union, non-union fractures and osteoporosis.
Indications d'application Optimal working dilution should be determined by the investigator.
Protocole This ELISA Pair Set is a solid phase sandwich ELISA (Enzyme-Linked Immunosorbent Assay). It utilizes a monoclonal antibody specific for BMP-2 coated on a 96-well plate. Standards and samples are added to the wells, and any BMP-2 present binds to the immobilized antibody. The wells are washed and a horseradish peroxidase conjugated mouse anti-BMP-2 monoclonal antibody is then added, producing an antibody-antigen-antibody "sandwich". The wells are again washed and TMB substrate solution is loaded, which produces color in proportion to the amount of BMP-2 present in the sample. To end the enzyme reaction, the stop solution is added and absorbances of the microwell are read at 450 nm.
Préparation des réactifs
  1. Dilute the capture antibody to the working concentration in CBS. Immediately coat a 96-well microplate with 100 μL per well of the diluted capture antibody. Seal the plate and incubate overnight at 4 °C.
    2. Aspirate each well and wash with at least 300 μL wash buffer, repeating the process two times for a total of three washes. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining wash buffer by inverting the plate and blotting it against clean paper towels.
    3. Block plates by adding 300 μL of blocking buffer to each well. Incubate at room temperature for a minimum of 1 hour.
    4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Procédure de l'essai
  1. Add 100 μL of sample or standards in sample dilution buffer per well. Seal the plate and incubate 2 hours at roomtemperature.
    2. Repeat the aspiration/wash as in step 2 of plate preparation.
    3. Add 100 μL of the detection antibody, diluted in antibody dilution buffer, to each well. Seal the plate and incubate 1hour at room temperature.
    4. Repeat the aspiration/wash as in step 2 of plate preparation.
    5. Add 200 μL of substrate solution to each well. Incubate for 20 minutes at room temperature (if substrate solutionis not as requested, the incubation time should be optimized). Avoid placing the plate in direct light.
    6. Add 50 μL of stop solution to each well. Gently tap the plate to ensure thorough mixing.
    7. Determine the optical density of each well immediately, using a microplate reader set to 450 nm.
Calcul des résultats

Calculate the mean absorbance for each set of duplicate standards, controls and samples. Subtract the mean zero standard absorbance from each. Construct a standard curve by plotting the mean absorbance for each standard on the y-axis against the concentration on the x-axis and draw a best fit curve through the points on the graph. To determine the concentration of the unknowns, find the unknowns' mean absorbance value on the y-axis and draw a horizontal line to the standard curve. At the point of intersection, draw a vertical line to the x-axis and read the concentration. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor. Alternatively, computer-based curve-fitting statistical software may also be employed to calculate the concentration of the sample.

Restrictions For Research Use only
Format Liquid
Conseil sur la manipulation Avoid repeated freeze-thaw cycles.
Stock -20 °C,-80 °C
Stockage commentaire Capture Antibody: Aliquot and store at -20°C to -80°C for up to 6 months from date of receipt. Avoid repeated freeze-thaw cycles.

Detection Antibody: Protect it from prolonged exposure to light. Aliquot and store at -20°C to -80°C and for up to 6 months from date of receipt. Avoid repeated freeze-thaw cycles.

Standard: Store lyophilized standard at -20°C to -80°C for up to 6 months from date of receipt. Aliquot and store the reconstituted Standard at -80°C for up to 1 month. Avoid repeated freeze-thaw cycles.
Date de péremption 6 months
Images (Fournisseur)
 image for Human BMP-2 ELISA Pair Set (ABIN2010205) Human BMP-2 ELISA Pair Set
Produit citée dans: Fu, Zhang, Sun, Liao, Bai, Zhang, Du, Jin, Wang, Li, Wang: "Controlled-release of bone morphogenetic protein-2 from a microsphere coating applied to acid-etched Ti6AL4V implants increases biological bone growth in vivo." dans: Journal of orthopaedic research : official publication of the Orthopaedic Research Society, Vol. 32, Issue 6, pp. 744-51, 2014 (PubMed).