PKC alpha ELISA Kit

Catalog No. ABIN5658216

This Colorimetric ELISA kit is designed for the quantitative measurement of Rat PKC alpha.

Quick Overview for PKC alpha ELISA Kit (ABIN5658216)

Target: PKC alpha (PKCa) (Protein Kinase C, alpha (PKCa))

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

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

Application: ELISA

Sample Type: Cell Lysate, Tissue Homogenate

Product Details PKC alpha ELISA Kit

Minimum Detection Limit: 0.312 ng/mL

Purpose: Rat Protein Kinase C Alpha (PKCa) ELISA Kit for detecting PKCa in tissue homogenates, cell lysates

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of Protein Kinase C Alpha (PKCa). No significant cross-reactivity or interference between Protein Kinase C Alpha (PKCa) and analogues was observed.

Sensitivity: 0.117 ng/mL

Components:

• Pre-coated, ready to use 96-well strip plate
• Standar
• Detection Reagent A
• Detection Reagent B
• TMB Substrate
• Stop Solution
• Assay Diluent A
• Assay Diluent B
• Wash Buffer (30x concentrate)
• Standard Diluent
• Plate Sealer
• Instruction Manual

Material not included:

• Microplate Reader with 450 nm filter
• Single or multi-channel pipettes with high precision and disposable tips
• Microcentrifuge tubes
• Deionized or distilled watter
• Absorbent paper for blotting the microplate
• Container for wash solution
• 0.01 M (or 1x) Phosphate Buffered Saline (PBS), ph 7.0-7.2

Application Details

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

Comment:

This product is tested and assembled before shipping to ensure that it meets its quality specifications. All kits are tested to confirm that they fall within their defined Inter- and Intra- assay coefficient of variation.

Sample Volume: 100 μL

Assay Time: 3 h

Plate: Pre-coated

Protocol: 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 Protein Kinase C Alpha (PKCa). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Protein Kinase C Alpha (PKCa). 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 Protein Kinase C Alpha (PKCa), 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 Protein Kinase C Alpha (PKCa) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Assay Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Protein Kinase C Alpha (PKCa) were tested 20 times on one plate, respectively
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Protein Kinase C Alpha (PKCa) 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

Handling

Handling Advice: The Stop Solution is acidic. Do not allow to contact skin or eyes. Calibrators, controls and specimen samples should be assayed in duplicate. Once the procedure has been started, all steps should be completed without interruption.

Storage: 4 °C,-20 °C

Storage Comment: -20°C. Bring all reagents to room temperature before beginning test. The kit may be stored at 4°C for immediate use within two days upon arrival. Reseal any unused strips with desiccant pack. Minimize freeze/thaw cycles.

Expiry Date: 12 months

Target Details for PKC alpha

Target: PKC alpha (PKCa) (Protein Kinase C, alpha (PKCa))

Alternative Name: Protein Kinase C Alpha

Background: Gene Aliases: PRKCA, PKC-A, AAG6, PKC-Alpha

UniProt: P05696

Pathways: WNT Signaling, TCR Signaling, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Thyroid Hormone Synthesis, cAMP Metabolic Process, Myometrial Relaxation and Contraction, Cell-Cell Junction Organization, Regulation of G-Protein Coupled Receptor Protein Signaling, G-protein mediated Events, Signaling Events mediated by VEGFR1 and VEGFR2, Interaction of EGFR with phospholipase C-gamma, Thromboxane A2 Receptor Signaling, VEGFR1 Specific Signals, VEGF Signaling