AKT1 ELISA Kit

Catalog No. ABIN6973778

Product Details AKT1 ELISA Kit

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

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.312 ng/mL - 20 ng/mL

Minimum Detection Limit: 0.312 ng/mL

Application: ELISA

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

Sample Type: Cell Lysate, Plasma, Serum

Analytical Method: Quantitative

Specificity: 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.

Sensitivity: 0.078 ng/mL

Components:

• 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

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Sample Volume: 100 μL

Assay Time: 1 - 4.5 h

Plate: Pre-coated

Protocol:

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.

Reagent Preparation:

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.

Sample Preparation:

• 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).

Assay Precision: 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

Handling

Storage: 4 °C,-20 °C

Storage Comment: 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.

Expiry Date: 6 months

Target Details for AKT1

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

Alternative Name: v-akt murine thymoma viral oncogene homolog 1 (AKT1 Products)

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

Background:

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: PI3K-Akt Signaling, RTK Signaling, TCR Signaling, AMPK Signaling, Interferon-gamma Pathway, TLR Signaling, 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, Warburg Effect