AKT1 Kit ELISA

N° du produit ABIN6960409

Détail du produit AKT1 Kit ELISA

Antigène: AKT1 (V-Akt Murine Thymoma Viral Oncogene Homolog 1 (AKT1))

Reactivité: Souris

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 0.15 ng/mL - 10 ng/mL

Seuil minimal de détection: 0.15 ng/mL

Application: ELISA

Fonction: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of PKBa in mouse tissue homogenates, cell lysates.

Type d'échantillon: Cell Lysate, Tissue Homogenate

Analytical Method: Quantitative

Specificité: This assay has high sensitivity and excellent specificity for detection of Protein Kinase B Alpha (PKBa)

Sensibilité: 0.055 ng/mL

Ingrédients:

• 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

Information d'application

Commentaires:

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.

Volume d'échantillon: 100 μL

Durée du test: 3 h

Plaque: Pre-coated

Protocole:

1. Prepare all reagents, samples and standards,
2. Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
4. Aspirate and wash 3 times,
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
6. Aspirate and wash 5 times,
7. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
8. Add 50μL Stop Solution. Read at 450nm immediately.

Préparation des réactifs:

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 20 ng/mL. Firstly dilute the stock solution to 10 ng/mL and the diluted standard serves as the highest standard (10 ng/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 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.312 ng/mL, 0.156 ng/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0 ng/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.

Préparation de l'échantillon:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Précision du teste: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target 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

Stockage

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.

Stock: 4 °C/-20 °C

Stockage commentaire:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

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Date de péremption: 6 months

Références pour AKT1 Kit ELISA (ABIN6960409)

Campo, Avenoso, DAscola, Scuruchi, Calatroni, Campo: "Beta-arrestin 1 is involved in the catabolic response stimulated by hyaluronan degradation in mouse chondrocytes." dans: Cell and tissue research, Vol. 361, Issue 2, pp. 567-79, (2015) (PubMed).

Détail du antigène

Antigène: AKT1 (V-Akt Murine Thymoma Viral Oncogene Homolog 1 (AKT1))

Autre désignation: Protein Kinase B Alpha (PKBa) (AKT1 Produits)

Synonymes: AKT Kit ELISA, CWS6 Kit ELISA, PKB Kit ELISA, PKB-ALPHA Kit ELISA, PRKBA Kit ELISA, RAC Kit ELISA, RAC-ALPHA Kit ELISA, Akt Kit ELISA, Rac Kit ELISA, PKB/Akt Kit ELISA, PKBalpha Kit ELISA, AKT/PKB Kit ELISA, AKT1 Kit ELISA, Akt/PKB Kit ELISA, Akt1 Kit ELISA, CG4006 Kit ELISA, D-Akt Kit ELISA, DAKT1 Kit ELISA, DAKT1/PKB Kit ELISA, DAkt Kit ELISA, DAkt1 Kit ELISA, DPKB Kit ELISA, DRAC-PK Kit ELISA, DRAC-PK66 Kit ELISA, DRAC-PK85 Kit ELISA, Dakt Kit ELISA, Dakt1 Kit ELISA, Dmel\\CG4006 Kit ELISA, Dpkb Kit ELISA, PKB/AKT Kit ELISA, PKB/dAKT Kit ELISA, RacPK Kit ELISA, akt Kit ELISA, akt1 Kit ELISA, dAKT Kit ELISA, dAKT/dPKB Kit ELISA, dAKT1 Kit ELISA, dAkt Kit ELISA, dAkt/PKB Kit ELISA, dAkt1 Kit ELISA, dPKB Kit ELISA, dakt Kit ELISA, dakt1 Kit ELISA, l(3)04226 Kit ELISA, l(3)89Bq Kit ELISA, p-Akt Kit ELISA, pAkt Kit ELISA, ACT-5 Kit ELISA, akt-1 Kit ELISA, pkb Kit ELISA, v-akt Kit ELISA, v-akt1 Kit ELISA, xAct Kit ELISA, ATAKT1 Kit ELISA, F18A8.2 Kit ELISA, F18A8_2 Kit ELISA, K+ transporter 1 Kit ELISA, POTASSIUM TRANSPORTER Kit ELISA, AKT serine/threonine kinase 1 Kit ELISA, thymoma viral proto-oncogene 1 Kit ELISA, CG4006 gene product from transcript CG4006-RE Kit ELISA, actin beta Kit ELISA, v-akt murine thymoma viral oncogene homolog 1 S homeolog Kit ELISA, v-akt murine thymoma viral oncogene homolog 1 Kit ELISA, K+ transporter 1 Kit ELISA, Serine/threonine-protein kinase akt-1 Kit ELISA, AKT1 Kit ELISA, Akt1 Kit ELISA, ACTB Kit ELISA, akt1.S Kit ELISA, akt1 Kit ELISA, KT1 Kit ELISA, akt-1 Kit ELISA

Pathways: Signalisation PI3K-Akt, Signalisation RTK, TCR Signaling, AMPK Signaling, Interferon-gamma Pathway, Signalisation TLR, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Response to Water Deprivation, Regulation of Actin Filament Polymerization, Carbohydrate Homeostasis, Glycosaminoglycan Metabolic Process, Cellular Glucan Metabolic Process, Regulation of Muscle Cell Differentiation, Cell-Cell Junction Organization, Regulation of Cell Size, Skeletal Muscle Fiber Development, Regulation of Carbohydrate Metabolic Process, Hepatitis C, Protein targeting to Nucleus, CXCR4-mediated Signaling Events, Signaling Events mediated by VEGFR1 and VEGFR2, Negative Regulation of intrinsic apoptotic Signaling, Thromboxane A2 Receptor Signaling, Signaling of Hepatocyte Growth Factor Receptor, Positive Regulation of fat Cell Differentiation, VEGFR1 Specific Signals, VEGF Signaling, L'effet Warburg