AKT1 Kit ELISA

N° du produit ABIN6973778

Détail du produit AKT1 Kit ELISA

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

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 0.312 ng/mL - 20 ng/mL

Seuil minimal de détection: 0.312 ng/mL

Application: ELISA

Fonction: For the quantitative determination of human RAC-alpha serine/threonine- protein kinase (AKT1) concentrations in serum, plasma, cell lysates.

Type d'échantillon: Cell Lysate, Plasma, Serum

Analytical Method: Quantitative

Specificité: This assay has high sensitivity and excellent specificity for detection of human AKT1. No significant cross-reactivity or interference between human AKT1 and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between human AKT1 and all the analogues, therefore, cross reaction may still exist.

Sensibilité: 0.078 ng/mL

Ingrédients:

• Assay plate
• Standard
• HRP-avidin (100 x concentrate)
• Biotin-antibody (100 x concentrate)
• Sample Diluent
• HRP-avidin Diluent
• Biotin-antibody Diluent
• Wash Buffer (25 x concentrate)
• TMB Substrate
• Stop Solution
• Adhesive Strip

Information d'application

Indications d'application: Optimal working dilution should be determined by the investigator.

Volume d'échantillon: 100 μL

Durée du test: 1 - 4.5 h

Plaque: Pre-coated

Protocole:

1. Prepare reagents, samples and standards as instructed.
2. Add 100 µL standard or sample to each well. Incubate 2 hours at 37 °C.
3. Remove the liquid of each well, don't wash.
4. Add 100 µL Biotin-antibody (1x) to each well. Incubate 1 hour at 37 °C.
5. Aspirate and wash 3 times.
6. Add 100 µL HRP-avidin (1x) to each well. Incubate 1 hour at 37 °C
7. Aspirate and wash 5 times.
8. Add 90 μL of TMB Substrate to each well. Incubate for 15-30 minutes at 37 °C. Protect from light.
9. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.

Préparation des réactifs:

1. Biotin-antibody (1x) - Centrifuge the vial before opening. Biotin-antibody requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of Biotin-antibody + 990 μL of Biotin-antibody Diluent.
2. HRP-avidin (1x) - Centrifuge the vial before opening. HRP-avidin requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-avidin + 990 μL of HRP-avidin Diluent.
3. Wash Buffer (1x) - If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 mL of Wash Buffer (1 x).
4. Standard Centrifuge the standard vial at 6000-10000rpm for 30s. Reconstitute the Standard with 1.0 mL of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution of 20 ng/mL. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions. Pipette 250 μL of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 2-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (20 ng/mL). Sample Diluent serves as the zero standard (0 ng/mL).

Note:

• Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit.
• Bring all reagents to room temperature (18-25 °C) before use for 30 min.
• Prepare fresh standard for each assay. Use within 4 hours and discard after use.
• Making serial dilution in the wells directly is not permitted.
• Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10 μL for once pipetting.
• Distilled water is recommended to be used to make the preparation for reagents. 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): CV%<8% Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10% Three samples of known concentration were tested in twenty assays to assess.

Restrictions: For Research Use only

Stockage

Stock: 4 °C,-20 °C

Stockage commentaire: Unopened kit Store at 2 - 8°C. Do not use the kit beyond the expiration date. May be stored for up to 1 month at 2 - 8°C. Coated assay Try to keep it in a sealed aluminum foil bag, plate and avoid the damp. Standard May be stored for up to 1 month at 2 - 8° C. If Biotin-antibody don't make recent use, better keep it store at HRP-avidin -20°C. Biotin-antibody Diluent Opened kit HRP-avidin Diluent Sample May be stored for up to 1 month at 2 - 8°C. Diluent Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.

Date de péremption: 6 months

Détail du antigène

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

Autre désignation: v-akt murine thymoma viral oncogene homolog 1 (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

Sujet:

Abbreviation: AKT1

Alias: AKT, MGC99656, PKB, PKB-ALPHA, PRKBA, RAC, RAC-ALPHA, AKT1 kinase|RAC-alpha serine/threonine-protein kinase|protein kinase B|rac protein kinase alpha

UniProt: P31749

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