ARG Kit ELISA
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- Antigène Voir toutes ARG Kits ELISA
- ARG (Arginase (ARG))
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Reactivité
- Souris
- Méthode de détection
- Colorimetric
- Type de méthode
- Sandwich ELISA
- Gamme de detection
- 15.6 pg/mL - 1000 pg/mL
- Seuil minimal de détection
- 15.6 pg/mL
- Application
- ELISA
- Type d'échantillon
- Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
- Analytical Method
- Quantitative
- Specificité
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This assay has high sensitivity and excellent specificity for detection of Mini Samples Arginase (ARG).
No significant cross-reactivity or interference between Mini Samples Arginase (ARG) and analogues was observed. - Sensibilité
- 6.4 pg/mL
- Classe de qualité
- Small Sample
- Ingrédients
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- Pre-coated, ready to use 96-well strip plate
- Plate sealer for 96 wells
- Standard Diluent
- Assay Diluent A
- Assay Diluent B
- Stop Solution
- Standard
- Detection Reagent A
- Detection Reagent B
- TMB Substrate
- Wash Buffer (30 x concentrate)
- Instruction manual
- Matériel non inclus
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- Microplate reader with 450 nm filter.
- Precision single or multi-channel pipettes and disposable tips.
- Eppendorf Tubes for diluting samples.
- Deionized or distilled water.
- Absorbent paper for blotting the microtiter plate.
- Container for Wash Solution
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- Durée du test
- 3 h
- Plaque
- Pre-coated
- Protocole
- 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 Mini Samples Arginase (ARG). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Mini Samples Arginase (ARG). 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 Mini Samples Arginase (ARG), 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 Mini Samples Arginase (ARG) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
- Préparation des réactifs
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- 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.
- 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 1,000pg/mL. Prepare 7 tubes containing 0.5 mL Standard Diluent and produce a double dilution series by transferring 500 µL each. 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.
- 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 stockDetection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
- Wash Solution - Dilute 10 mL of Wash Solution concentrate (30x) with 290 mL of deionized or distilled water to prepare 300 mL of Wash Solution (1x).
- TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.
Note:
- Making serial dilution in the wells directly is not permitted.
- Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
- 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 once pipetting.
- The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
- Prepare Substrate working Solution within 15 minutes before assay.
- If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
- Contaminated water or container for reagent preparation will influence the detection result.
- Procédure de l'essai
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- Prepare all reagents, samples and standards,
- Add 25μL standard or sample to each well. Incubate 1 hour at 37 °C,
- Aspirate and add 25μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
- Aspirate and wash 3 times,
- Add 25μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
- Aspirate and wash 5 times,
- Add 25μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
- Add 20μL Stop Solution. Read at 450nm immediately.
- Prepare all reagents, samples and standards,
- Précision du teste
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Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mini Samples Arginase (ARG) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mini Samples Arginase (ARG) 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
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- Précaution d'utilisation
- The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
- Conseil sur la manipulation
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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. - Stock
- 4 °C
- Stockage commentaire
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- 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.
- Date de péremption
- 6 months
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- Antigène Voir toutes ARG Kits ELISA
- ARG (Arginase (ARG))
- Abstract
- ARG Produits
- Synonymes
- CG18104 Kit ELISA, Dmel\\CG18104 Kit ELISA, EG:171D11.4 Kit ELISA, EG:65F1.3 Kit ELISA, SI:zC146F4.4 (novel protein with NUDIX domain) Kit ELISA, si:ch211-146f4.3 Kit ELISA, Tb08.26N11.490 Kit ELISA, rocF Kit ELISA, NV10213 Kit ELISA, argi1 Kit ELISA, arginase Kit ELISA, arginase 1 L homeolog Kit ELISA, arginase 1 Kit ELISA, Arginase-1 Kit ELISA, arg Kit ELISA, arg1.L Kit ELISA, arg1 Kit ELISA, ARGAH1 Kit ELISA, BP0538 Kit ELISA, rocF Kit ELISA, SAS2066 Kit ELISA, RR_RS05730 Kit ELISA, CND03500 Kit ELISA, CNG00550 Kit ELISA, Tb927.8.2020 Kit ELISA, PGTG_16455 Kit ELISA, LOC100123155 Kit ELISA, argi1 Kit ELISA
- UniProt
- Q61176
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