AKT1 ELISA Kit

Catalog No. ABIN7672471

This Rat AKT1 ELISA Kit is a Colorimetric ELISA Kit designed to quantify Rat AKT1.

Quick Overview for AKT1 ELISA Kit (ABIN7672471)

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

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.156 ng/mL - 10 ng/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Lysate

Grade: QuickTest

Product Details AKT1 ELISA Kit

Minimum Detection Limit: 0.156 ng/mL

Purpose: Rat AKT1 (RAC-alpha serine/threonine-protein kinase) QuickTest ELISA Kit

Brand: QuickTest

Analytical Method: Quantitative

Sensitivity: 0.094 ng/mL

Components:

• ELISA Microplate (Dismountable)
• Lyophilized Standard
• Capture/Detection Antibody
• HRP-Streptavidin (orange)
• TMB Substrate
• Sample Dilution Buffer (blue)
• Stop Solution
• Wash Buffer (25x)
• Plate Sealer
• Product Description

Material not included:

• Microplate reader (wavelength: 450nm)
• 37 °C incubator (CO2 incubator for cell culture is not recommenced.)
• Automated plate washer or multi-channel pipette/5 mL pipettor (for manual washing purpose)
• Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips (Calibration is required before use.)
• Sterile tubes and sample tubes with disposable tips
• Absorbent paper and loading slot
• Deionized or distilled water

Application Details

Comment:

Traditional sandwich ELISA assay usually takes 3-4 hours. Most detection reagents in the market are concentrated and can't be distinguished from the colour. Dilution is required before use. We developed QuickTest ELISA kits to decrease customers’ operational errors, improve user's experience and get accurate assay result more efficiently. Compared with regular ELISA kits, QuickTest ELISA kits have more advantages in the same linearity, stability, recovery and storage conditions.

Sample Volume: 50 µL

Assay Time: 2 h

Plate: Pre-coated

Protocol: Capture antibody was conjugated to an affinity tag that was recognized by a specific antibody pre-coated on the QuickTest plate. Add the Cap/Det Ab working solution into each well, then add the standards and pilot samples into relevant wells. Mix thoroughly and then incubate. If the sample contains detection target, a capture antibody-detection target-biotin-detection antibody complex was formed. After washing off unbound conjugates, HRP-Streptavidin was added to incubate. Repeat the washing process and then TMB substrates were added to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm with a microplate reader. The concentration of the detection target in the sample was calculated by plotting a standard curve. The concentration of the target substance is proportional to the OD450 value.

Assay Procedure:

1. Take out the required strips, add 50 µL Cap/Det Ab into each well, then add 50 µL Standard or Sample into relevant well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure complete mixing then statically incubate for 60 minutes at 37 °C.
2. Wash the plate twice without immersion.
3. Add 100 µL HRP-Streptavidin (orange) into each well, seal the plate and statically incubate for 30 minutes at 37 °C.
4. Wash the plate five times without immersion.
5. Add 90 µL TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37 °C (Accurate TMB visualization control is required.).
6. Add 50 µL stop solution. Read at 450nm immediately and calculate.

Restrictions: For Research Use only

Handling

Storage: 4 °C

Storage Comment: 2-8 °C for 12 months

Expiry Date: 12 months

Target Details for AKT1

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

Alternative Name: AKT1

Background: RAC-alpha serine/threonine-protein kinase|Protein kinase B|PKB|Protein kinase B alpha|PKB alpha|Proto-oncogene c-Akt|RAC-PK-alpha|AKT1|PKB|RAC

UniProt: P47196

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