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Osteocalcin ELISA Kit

BGLAP Reactivity: Rat Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Lysate, Plasma, Serum, Tissue Homogenate
Catalog No. ABIN416281
  • Target See all Osteocalcin (BGLAP) ELISA Kits
    Osteocalcin (BGLAP)
    Reactivity
    • 6
    • 5
    • 4
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    Rat
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Detection Range
    15.6 pg/mL - 1000 pg/mL
    Minimum Detection Limit
    15.6 pg/mL
    Application
    ELISA
    Purpose
    The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of OC in Serum,Plasma,Tissue Homogenate,Cell Lysate,Biological Fluids
    Sample Type
    Cell Lysate, Plasma, Serum, Tissue Homogenate
    Analytical Method
    Quantitative
    Specificity

    This assay has high sensitivity and excellent specificity for detection of Osteocalcin (OC).
    No significant cross-reactivity or interference between Osteocalcin (OC) and analogues was observed.

    Cross-Reactivity (Details)
    No significant cross-reactivity or interference between Osteocalcin (OC) and analogues was observed.
    Sensitivity
    6.3 pg/mL
    Components
    • 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
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  • Application Notes
    • Limited by the current condition and scientific technology, we cannot completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.
    • The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.
    • Kits from different batches may be a little different in detection range, sensitivity and color developing time.
    • Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
    • Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.
    • There may be some foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
    • Wrong operations during the reagents preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
    • Even the same operator might get different results in two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before assay for each batch is recommended.
    • Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.
    • Kits from different manufacturers for the same item might produce different results, since we have not compared our products with other manufacturers.
    Comment

    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.

    Sample Volume
    100 μL
    Assay Time
    3 h
    Plate
    Pre-coated
    Protocol
    The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Osteocalcin (OC). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Osteocalcin (OC). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Osteocalcin (OC), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Osteocalcin (OC) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
    Reagent Preparation
    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 5,000pg/mL. Firstly dilute the stock solution to 1,000pg/mL and the diluted standard serves as the highest standard (1,000pg/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 1,000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL, 15.6pg/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.
    Assay Precision

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

    Restrictions
    For Research Use only
  • Precaution of Use
    The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
    Handling Advice
    The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5 % within the expiration date under appropriate storage condition.
    To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
    Storage
    4 °C
    Storage Comment
    • For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
    • For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
      Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
    • For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C.
    Expiry Date
    6 months
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    Liu, Xiong, Wang, Yang, Wang, Liu, Wu, Li, Ou, Zhang, Zhu: "Effects of water extract from epimedium on neuropeptide signaling in an ovariectomized osteoporosis rat model." in: Journal of ethnopharmacology, Vol. 221, pp. 126-136, (2018) (PubMed).

    Chen, Xue, Shen, Mu, Fu: "Curcumin alleviates glucocorticoid-induced osteoporosis through the regulation of the Wnt signaling pathway." in: International journal of molecular medicine, Vol. 37, Issue 2, pp. 329-38, (2016) (PubMed).

    de Cássia Alves Nunes, Chiba, Pereira, Pereira, de Lima Coutinho Mattera, Ervolino, Louzada, Buzalaf, Silva, Sumida: "Effect of Sodium Fluoride on Bone Biomechanical and Histomorphometric Parameters and on Insulin Signaling and Insulin Sensitivity in Ovariectomized Rats." in: Biological trace element research, Vol. 173, Issue 1, pp. 144-53, (2016) (PubMed).

    Chen, Wang, Wang, Yin, Gao, Xiang, Liu, Xiong, Wang, Zhu, Yang, Zhang: "Antiosteoporotic effect of icariin in ovariectomized rats is mediated via the Wnt/β-catenin pathway." in: Experimental and therapeutic medicine, Vol. 12, Issue 1, pp. 279-287, (2016) (PubMed).

    Nowak, Matuszewska, Filipiak, Nikodem, Merwid-Ląd, Pieśniewska, Fereniec-Gołębiewska, Kwiatkowska, Szeląg: "The influence of bexarotene, a selective agonist of the retinoid receptor X (RXR), and tazarotene, a selective agonist of the retinoid acid receptor (RAR), on bone metabolism in rats." in: Advances in medical sciences, Vol. 61, Issue 1, pp. 85-9, (2016) (PubMed).

    Stringhetta-Garcia, Singulani, Santos, Louzada, Nakamune, Chaves-Neto, Rossi, Ervolino, Dornelles: "The effects of strength training and raloxifene on bone health in aging ovariectomized rats." in: Bone, Vol. 85, pp. 45-54, (2016) (PubMed).

    Zhang, Chen, Wei, Liu, Lei, Bai: "Ginsenosides Rg3 attenuates glucocorticoid-induced osteoporosis through regulating BMP-2/BMPR1A/Runx2 signaling pathway." in: Chemico-biological interactions, Vol. 256, pp. 188-97, (2016) (PubMed).

    Matuszewska, Nowak, Rzeszutko, Zduniak, Szandruk, Jędrzejuk, Landwójtowicz, Bolanowski, Pieśniewska, Kwiatkowska, Szeląg: "Effects of long-term administration of pantoprazole on bone mineral density in young male rats." in: Pharmacological reports : PR, Vol. 68, Issue 5, pp. 1060-4, (2016) (PubMed).

    Shalan, Mustapha, Mohamed: "Noni leaf and black tea enhance bone regeneration in estrogen-deficient rats." in: Nutrition (Burbank, Los Angeles County, Calif.), Vol. 33, pp. 42-51, (2016) (PubMed).

    Pereira, Chiba, de Lima Coutinho Mattera, Pereira, de Cássia Alves Nunes, Tsosura, Okamoto, Sumida: "Effects of fluoride on insulin signaling and bone metabolism in ovariectomized rats." in: Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements (GMS), Vol. 39, pp. 140-146, (2016) (PubMed).

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  • Target See all Osteocalcin (BGLAP) ELISA Kits
    Osteocalcin (BGLAP)
    Alternative Name
    Osteocalcin (BGLAP Products)
    Synonyms
    BGP ELISA Kit, OC ELISA Kit, OCN ELISA Kit, Bglap1 ELISA Kit, Bglap2 ELISA Kit, OG1 ELISA Kit, mOC-A ELISA Kit, Bgp ELISA Kit, Bgpr ELISA Kit, Bgpra ELISA Kit, bgp ELISA Kit, LOC792433 ELISA Kit, ocn ELISA Kit, GLA ELISA Kit, BGLAP ELISA Kit, PMF1 ELISA Kit, bone gamma-carboxyglutamate protein ELISA Kit, bone gamma carboxyglutamate protein ELISA Kit, bone gamma-carboxyglutamate protein S homeolog ELISA Kit, bone gamma-carboxyglutamate (gla) protein ELISA Kit, osteocalcin ELISA Kit, polyamine-modulated factor 1 ELISA Kit, BGLAP ELISA Kit, Bglap ELISA Kit, bglap.S ELISA Kit, bglap ELISA Kit, ocn ELISA Kit, LOC100529177 ELISA Kit, LOC100146589 ELISA Kit
    UniProt
    P04640
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