SKU: WT195E Category:

U-Blot® Ubiquitinated PTEN ELISA Kit

$999.00

SKU: WT195E
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Product details

The Ubiquitinated-PTEN and Total-PTEN ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human ubiquitinated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan PTEN antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and PTEN present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-ubiquitination antibody is used to detect ubiquitinated PTEN or biotinylated anti-PTEN antibody is used to detect total PTEN . After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of ubiquitinated PTEN or total PTEN bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.This assay enables quick and easy quantification of target proteins and allows high-throughput screening of numerous different lysates without the need for traditional Western Blot analysis. It features minimal hands-on operation time, simple workflow, and completely non-radioactive detection system, ensuring safe and convenient experimental operation.

Specifications

Size1 Plate Kit, 2 Plate Kit, 5 Plate Kit
SpeciesHuman
Compatible Sample TypesTissue Lysates, Cell Lysates
Accession NumberP60484
Gene Id5728
Gene SymbolsPTEN, MMAC1, TEP1
Protein Name / SynonymsPhosphatidylinositol 3,4,5-trisphosphate 3-phosphatase and dual-specificity protein phosphatase PTEN; Inositol polyphosphate 3-phosphatase; Mutated in multiple advanced cancers 1; Phosphatase and tensin homolog
Solid Support96-well Microplate
Design PrincipleSandwich-based
Method Of DetectionColorimetric
Research AreaPost-Translational Modifications, Epigenetics, Akt Signaling
ModificationUbiquitination
Shipping Type2‑8 ℃
Storage-20°C
Kit ComponentsMicroplate Coated With Anti-Pan-PTEN Antibody:1 plate;Anti-Ubiquitination Detection Antibody:1 vial;Anti-PTEN Detection Antibody:1 vial;HRP-Conjugated Streptavidin:200 µL;Wash Buffer:25 mL;Assay Diluent:15 mL;Lysis Buffer:5 mL;TMB One-Step Substrate Reagent:12 mL;Stop Solution:8 mL;
Additional Materials RequiredMicroplate reader capable of measuring absorbance at 450 nm;Protease and Phosphatase inhibitors;Precision pipettes to deliver 2 µL to 1 mL volumes;Adjustable 1-25 mL pipettes for reagent preparation;100 mL and 1 liter graduated cylinders;Log-log graph paper or computer and software for ELISA data analysis;Absorbent paper;Tubes to prepare sample dilutions;Distilled or deionized water;Shaker
Assay Procedure SummaryPrepare all reagents and samples as instructed; Add 100 µL sample to each well. Incubate 2.5 hours at room temperature or overnight at 4°C with gentle shaking; Add 100 µL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking; Add 100 µL prepared HRP-Conjugated Streptavidin solution. Incubate for 1 hour at room temperature with gentle shaking; Add 100 µL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature; Add 50 µL Stop Solution to each well. Read at 450 nm immediately.
Storage/StabilityThe entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.
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