UMP/CMP-Glo™ Glycosyltransferase Assay
Universal Glycosyltransferase Activity Assay
- Easy, "add-incubate-read" protocol
- Detects sialyltransferases and phosphoglycosyltransferases that use CMP-, CDP- or UDP-sugars as donor substrates
- Ideal for low-activity glycosyltransferases
Catalog Number:
Size
Catalog Number: VA1130
Catalog Number: VA1131
Catalog Number: VA1132
シンプルな糖転移酵素発光アッセイ
UMP/CMP-Glo™ Glycosyltransferase Assay はグリコシルトランスフェラーゼ反応で UMP や CMP 産生を迅速に検出するホモジニアスな1液タイプのアッセイです。グリコシルトランスフェラーゼ反応後に等量の UMP/CMP Detection Reagent を加えると同時に生成されたUMP または CMP がATPに変換され、ルシフェラーゼ反応で光を生じます。発光はルミノメーターで検出し、発光値は nMレベルから50μMの UMP または CMP の濃度に比例します。UMP または CMP 標準曲線により発光値からUMP または CMP の濃度を求めることができます。
Add-Mix-Read for Results
Start with purified enzymes, sugar donors and acceptor molecules, and you can detect the reaction products UMP or CMP from phosphoglycosyltransferases or sialyltransferases, respectively. Just add the UMP/CMP Detection Reagent and measure luminescence.
Works with Many Sialyltransferases and Phosphoglycosyltransferases
The UMP/CMP-Glo™ Assay can be used to measure the activity of just about any sialyltransferase that uses CMP-sugars or any phosphoglycosyltransferase that uses UDP-sugars as a substrate. Here, we looked at sialyltransferases involved in protein O-glycosylation.
Ideal for Low-Activity Glycosyltransferases and Phosphoglycosyltransferases
The sensitivity of the UMP/CMP-Glo™ Glycosyltransferase Assay means it can detect low UMP or CMP concentrations with high dynamic range. Therefore, whether assaying for a low-activity glycosyltransferase whose sugar transfer rate is low or using a low concentration of enzyme that produces a small amount of UMP or CMP, you can obtain a high signal-to-background ratio using the assay.
The data below show the linearity and sensitivity of the UMP/CMP-Glo™ Glycosyltransferase Assay. Here, a UMP or CMP standard curve was prepared over the indicated concentration range in 25µl of 1X glycosyltransferase reaction buffer in a solid white 96-well plate. UMP/CMP-Glo™ Glycosyltransferase Assay was performed using 25µl of UMP/CMP Detection Reagent at room temperature. Luminescence was recorded using a GloMax® luminometer. Values represent the mean of two replicates. The tables show luminescence measured 1 hour after adding the UMP/ CMP Detection Reagent, and signal-to-background ratios calculated for each concentration of the standard curve.
Luminescent Signal is Stable Over Several Hours
The UMP/CMP-Glo™ Assay generates a highly stable luminescent signal that is stable for more than 3 hours, making it convenient to batch-process assay plates.
The data here show that the signal obtained with the UMP/CMP-Glo™ Assay is stable for several hours. To determine signal stability, luminescence was recorded at 1 and 3 hours after adding detection reagent. The signal-to-background ratio did not change over the time measured (data not shown). RLU = relative light units.
Protocols
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
ATP Detection Substrate |
V363C | 1 × 1 vial |
Nucleotide Detection Buffer |
V696A | 1 × 5ml |
UMP, 10mM |
VA113A | 1 × 100μl |
CMP, 10mM |
VA114A | 1 × 100μl |
UMP/CMP-Glo™ Enzyme |
VA115A | 1 × 200μl |
SDS
Search for SDS分析証明書
使用制限
For Research Use Only. Not for Use in Diagnostic Procedures.保存条件
See Protocol for detailed storage recommendations.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
|
ATP Detection Substrate |
V363A | 1 × 1 vial |
Nucleotide Detection Buffer |
V696B | 1 × 10ml |
UMP, 10mM |
VA113A | 1 × 100μl |
CMP, 10mM |
VA114A | 1 × 100μl |
UMP/CMP-Glo™ Enzyme |
VA115B | 1 × 400μl |
SDS
Search for SDS分析証明書
使用制限
For Research Use Only. Not for Use in Diagnostic Procedures.保存条件
See Protocol for detailed storage recommendations.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
|
ATP Detection Substrate |
V363B | 1 × 1 vial |
Nucleotide Detection Buffer |
V696C | 1 × 100ml |
UMP, 10mM |
VA113A | 1 × 100μl |
CMP, 10mM |
VA114A | 1 × 100μl |
UMP/CMP-Glo™ Enzyme |
VA115C | 4 × 1ml |
SDS
Search for SDS分析証明書
使用制限
For Research Use Only. Not for Use in Diagnostic Procedures.保存条件
See Protocol for detailed storage recommendations.
Related Products
類似製品
UDP-Glo™ Glycosyltransferase Assay
" 添加 - インキュベート - 測定 " だけの簡便でユニバーサルな糖転移酵素活性アッセイ
V6961, V6962, V6963, V6971, V6972, V6981, V6982, V6991, V6992, V7051, V7052, V7061, V7062
GDP-Glo™ Glycosyltransferase Assay
" 添加 - インキュベート - 測定 " だけの簡便でユニバーサルな糖転移酵素活性アッセイ
VA1090, VA1091, VA1092, VA1093, VA1094, VA1095, VA1096
関連製品
Ultra Pure UDP-Sugar Substrates
UDP-Glo™ Glycosyltransferase Assay とともに使用する基質
V7071, V7072, V7081, V7082, V7091, V7092, V7171, V7172, V7321, V7322
GloMax® Explorer System
幅広い測定に適応する高性能マルチモード検出装置
GM3500, GM3510
GloMax® Discover System
発光・蛍光・吸光度測定に対応した高性能マイクロプレートリーダー
GM3000
GloMax® Navigator System
使いやすいマイクロプレートルミノメーター
GM2000, GM2010
