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EpiQuik In Vivo Universal Protein Sumoylation Assay Kit

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Suggested Workflow
Nuclear Protein Extraction
 
 
SUMOylation
 
Input Type: Nuclear Extracts
Research Area: Sumoylation
Target Application: Amount Quantitation
Vessel Format: 96-Well Plate
100% Guarantee: 6 months
Catalog No.SizePriceQty
P-8001-4848 assays $338.00 
P-8001-9696 assays $549.00 
Availability: Usually Ships In 1-2 Days 
Product Overview

 

 

We recommend using a newer, improved version of this kit: EpiQuik In Vivo Protein Sumoylation Assay Ultra Kit (Colorimetric)

 

 

 

The EpiQuik™ In Vivo Universal Protein Sumoylation Assay Kit is a convenient set of tools that allows the experimenter to measure in vivo protein sumoylation. The kit is ready-to-use and provides all the essential components needed for measuring in vivo protein sumoylation from multiple mammalian cells/tissues including human, mouse and rat.

WHY CHOOSE THE EPIQUIK™ IN VIVO UNIVERSAL PROTEIN SUMOYLATION ASSAY KIT,

  • Fast procedure, which can be finished within 5 hours.
  • One-step colorimetric assay without the need for affinity chromatography and Western blotting.
  • Flexible antibody choice allows the detection of sumoylation of multiple target proteins simultaneously.
  • Included SUMO protein as the positive control allows protein sumoylation to be quantified.
  • Strip microplate format makes the assay flexible: manual or high throughput analysis.
  • Simple, reliable, and consistent assay conditions.
Product Details

The EpiQuik™ In Vivo Universal Protein Sumoylation Assay Kit is designed for measuring sumoylation of targeted proteins. Sumoylation of the targeted proteins is indicated by SUMO conjugated to these proteins. In an assay with this kit, the antibodies specific to the targeted proteins are stably bound to the strip wells and the targeted proteins are captured by these antibodies. Sumoylation of the targeted proteins is detected by recognition of SUMO conjugated to these proteins with an anti-SUMO antibody. The ratio or intensity of the sumoylation, which is proportional to the conjugated SUMO amount, can be quantified through the signal report-color development system.

SCHEMATIC PROCEDURE:  
 
Product Components

S1 (10X Wash Buffer)
S2 (Binding Buffer)
S3 (Mouse IgG, 1 µg/µl)*
S4 (Blocking Buffer)
S5 (Sumo Assay Buffer)
S6 (Sumo Protein, 1 µg/µl)*
S7 (Sumo Antibody, 1 µg/µl)*
S8 (Signal Report Solution)*
S9 (Color Development Solution)
S10 (Stop Solution)
8-Well Assay Strip (with Frame)
8-Well Control Strips
User Guide


* For maximum recovery of the products, centrifuge the original vial after thawing prior to opening the cap.

User Guide & MSDS

[User Guide]*
*Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

Customer Reviews

Rating By b*********@unmc.edu Verified Purchase Reviewed on: Tuesday 20 December, 2016
Application Description
Describe the experiment that the product used for. If an antibody, what application and protocol was used?
EpiQuik Invivo Universal protein Sumoylation Assay Kit, Cat No. P-8001

How friendly was the user guide? (1 – worst, 5 – best)
User guide is best, If treated and untreated sample can be define better that would be great

How professional was the appearance and presentation of the product? (1 – worst, 5 – best)
appearance and presentation of the product is best

How would you rate the overall product? (1 – Worst, 5 – best)
5-Best

Pros: Easy and faster to show in Vivo Sumoylation, Required small amount of protein

Cons: there was no cons of the product

Procedural Details
Sensitive Sumo-ELISA/Sandwich-ELISA assays validated that both extrinsic and intrinsic Prdx6 were Sumoylated, and showed that a fraction of endogeneous Prdx6 was present in Sumoylated form. (A) hLECs were transfected with pEGFP-vector or pEGFP-vector plus pEGFP-Sumo1 or pGFP-Prdx6 plus pEGFP-Sumo1. After 48h total cell lysates were prepared and submitted to Sandwich/Sumo1-ELISA assays to check the total Prdx6 protein and Sumoylated Prdx6 protein. Sumoylated Prdx6 protein was subtracted from total Prdx6 protein, presenting as Prdx6 unSumoylated (gray bars) and Sumoylated (black bars) forms. The data represent mean ± SD from three independent experiments (*p<0.001). (B) Prdx6+/+ LECs were transfected with pEGFP-vector plus pEGFP-Sumo1 or pGFP-Prdx6 plus pEGFP-Sumo1. Total cell lysates were prepared and used to perform Sandwich/Sumo1-ELISA. Sumoylated Prdx6 protein was subtracted from total Prdx6 protein, presenting as Prdx6 unSumoylated (gray bars) and Sumoylated (black bars) forms. The data represent the mean ± SD of three independent experiments (*p<0.001 vs control).

Other Thoughts
Overall kit is ok but company providing very less Sumoylation buffer. As now its known that Sumoylation occurs in nucleus as well as cytosol so total lysate or nuclear protein can be used. As many labs use own lysis buffer (RIPA) to isolate protein. Sometime protein concentration is low (eg 0.5ug/ul or 1ug/ul) so we have to increase the volume of protein so of course have to increase SUMO assay buffer. All buffer volume is ok but Sumoylation assay buffer volume is too low. Company can increase the volume of Sumoylation assay buffer or can provide the method.



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