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|Antigen||Caspase 12 (Gene/pseudogene) (CASP12) ELISA Kits|
Kits mit alternativen Reaktivitäten:
|Untere Nachweisgrenze||0.312 ng/mL|
|Hersteller||Anmelden zum Anzeigen|
Produktdetails Caspase 12 ELISA KitAntigendetails Anwendungsinformationen Handhabung Bilder
|Verwendungszweck||This immunoassay kit allows for the in vitro quantitative determination of human inactive caspase-12,CASP-12 concentrations in serum, Plasma, Urine, tissue homogenates and Cell culture supernates and Other biological fluids.|
|Proben||Plasma, Serum, Tissue Homogenate|
This assay recognizes recombinant and natural human CASP-12. No significant cross-reactivity or interference was observed.
Note: Limited by current skills and knowledge, it is impossible for us to complete the cross- reactivity detection between human CASP-12 and all the analogues, therefore, cross reaction may still exist.
|Sensitivität||< 0.12 ng/mL|
Assay plate x1
Sample Diluent 1 x 20ml
Assay Diluent A 1 x 10ml
Assay Diluent B 1 x 10ml
Detection Reagent A 1 x 120μl
Detection Reagent B 1 x 120μl
Wash Buffer(25 x concentrate) 1 x 30ml
Substrate 1 x 10ml
Stop Solution 1 x 10ml
Plate sealer for 96 wells x5
AntigendetailsProduktdetails Caspase 12 ELISA Kit Anwendungsinformationen Handhabung Bilder zurück nach oben
|Andere Bezeichnung||CASP12 (CASP12 ELISA Kit Abstract)|
|Hintergrund||Synonyms: Inactive caspase-12(CASP-12)|
|Pathways||Apoptose, ER-Nucleus Signaling, Unfolded protein response|
AnwendungsinformationenProduktdetails Caspase 12 ELISA Kit Antigendetails Handhabung Bilder zurück nach oben
|Testdauer||3 - 5 h|
|Protokoll||The microtiter plate provided in this kit has been pre-coated with an antibody specific to CASP-12. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for CASP-12 and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. Only those wells that contain CASP-12, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of CASP-12 in the samples is then determined by comparing the O.D. of the samples to the standard curve.|
|Aufbereitung der Reagenzien||
Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mix gently until the crystals have completely dissolved. Dilute 30 mL of Wash Buffer Concentrate into deionized or distilled water to prepare 750 mL of Wash Buffer.
Standard - Reconstitute the Standard with 1.0 mL of Sample Diluent. This reconstitution produces a stock solution of 20.0 ng/mL. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making serial dilutions (Making serial dilution in the wells directly is not permitted). The undiluted standard serves as the high standard (20.0 ng/mL). The Sample Diluent serves as the zero standard (0 ng/mL).
Detection Reagent A and B - Dilute to the working concentration using Assay Diluent A and B (1:100), respectively.
Serum - Use a serum separator tube (SST) and allow samples to clot for 30 minutes before centrifugation for 15 minutes at approximately 1000 x g. Remove serum and assay immediately or aliquot and store samples at -20 °C or -80 °C.
Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 x g at 2 °C - 8 °C within 30 minutes of collection. Store samples at -20 °C or -80 °C. Avoid repeated freeze-thaw cycles. Urine - Aseptically collect the first urine of the day (mid-stream), voided directly into a sterile container. Centrifuge to remove particulate matter, assay immediately or aliquot and store at ≤ -20 °C. Avoid repeated freeze-thaw cycles.
Tissue homogenates - The preparation of tissue homogenates will vary depending upon tissue type. For this assay, tissue was rinsed with 1X PBS to remove excess blood, homogenized in 20 mL of 1X PBS and stored overnight at ≤ -20 °C After two freeze-thaw cycles were performed to break the cell membranes, the homogenates were centrifuged for 5 minutes at 5000 x g. Remove the supernate and assay immediately or aliquot and store at ≤ -20 °C. Cell culture supernates and
Other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20 °C or -80 °C. Avoid repeated freeze-thaw cycles.
Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37 °C directly.). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at 4 °C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments.
Average the duplicate readings for each standard, control, and sample and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the CASP-12 concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. It is recommended to use some related software to do this calculation, such as curve expert 1.3. This procedure will produce an adequate but less precise fit of the data. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
|Beschränkungen||Nur für Forschungszwecke einsetzbar|
HandhabungProduktdetails Caspase 12 ELISA Kit Antigendetails Anwendungsinformationen Bilder zurück nach oben
|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.|
|Informationen zur Lagerung||
The Assay Plate, Standard, Detection Reagent A and Detection Reagent B should be stored at -20°C upon being received. After receiving the kit , Substrate should be always stored at 4°C.Other reagents are kept according to the labels on vials. But for long term storage, please keep the whole kit at -20°C. The unused strips should be kept in a sealed bag with the desiccant provided to minimize exposure to damp air. The test kit may be used throughout the expiration date of the kit (six months from the date of manufacture). Opened test kits will remain stable until the expiring date shown, provided it is stored as prescribed above.
Note: Because of the inherent stability of lyophilized materialthese materials may shipped at ambient temperature.