BCAA-Glo™ Assay
Easy, Sensitive Branched-Chain Amino Acid (Leu/Ile/Val) Assay Kit
- Compatible with cells, media, tissue and serum samples
- In-well sample preparation protocol streamlines workflow
- Scalable to 384-well plates for HTS applications
Catalog Number:
Size
Catalog Number: JE9300
Catalog Number: JE9400
Efficient BCAA Detection from Diverse Samples
The BCAA-Glo™ Assay is a fast and sensitive method for detecting branched-chain amino acids (BCAA; leucine, isoleucine and valine) from various biological samples, such as cultured cells, media, tissue homogenates and serum. Based on bioluminescent technology, the BCAA-Glo™ Assay can detect subtle changes in amino acid metabolism and abundance. Its streamlined sample preparation protocol involves acid treatment and neutralization directly in the assay wells, eliminating the need for cell collection, centrifugation and spin columns. The assay is adaptable to a 384-well format, facilitating quick analysis of many samples.
How the BCAA-Glo™ Assay Works
Leucine Dehydrogenase catalyzes the oxidation of BCAA, simultaneously reducing NAD+ to NADH. In the presence of NADH, Reductase converts the proluciferin Reductase Substrate into luciferin. This reaction is detected by Ultra-Glo™ Recombinant Luciferase, which emits light proportionate to the amount of BCAA in the sample.
Leucine Dehydrogenase recognizes all three BCAA with similar efficiency. It is specific for BCAA L-isomers.
Simple Add-and-Read Protocol Saves Time
Accurately Detect the Smallest Changes in Branched-Chain Amino Acids
|
Sensitivity (S/B >5) |
400nM |
|
|
Limit of Detection (S/N >3) |
50nM |
|
|
Linear Range |
50nM to 25μM |
|
|
Maximum Assay Window (S/B) |
>300 |
|
Sensitive Quantitation of BCAA in Cultured Cells
Cellular BCAA levels vary depending on cell type, growth conditions and metabolic activity. Intracellular BCAA concentrations are typically low. However, the BCAA-Glo™ Assay, with its high sensitivity and wide linear detection range, successfully measures BCAA levels in cell lysate samples ranging from 5,000 to 40,000 cells per 50μl, as shown in the graph.
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Protocols
Complete Protocol
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
Leucine Dehydrogenase |
J412A | 1 × 100μl | |
Leucine, 10mM |
J413A | 1 × 50μl | |
Reductase |
G884A | 1 × 55μl | 6mg/ml |
Reductase Substrate |
G885A | 1 × 55μl | |
Luciferin Detection Solution |
J135A | 1 × 5ml | |
NAD |
J136B | 1 × 275μl | |
Neutralization Buffer |
J153A | 1 × 15ml | |
NADP+ |
J154B | 1 × 100μl | |
0.6N HCl |
J402A | 1 × 15ml |
U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
Leucine Dehydrogenase |
J412B | 1 × 1ml | |
Leucine, 10mM |
J413A | 1 × 50μl | |
Reductase |
G884B | 1 × 275μl | 6mg/ml |
Reductase Substrate |
G885B | 1 × 275μl | |
Luciferin Detection Solution |
J135B | 1 × 50ml | |
NAD |
J136C | 1 × 1ml | |
Neutralization Buffer |
J153A | 1 × 15ml | |
NADP+ |
J154C | 1 × 500μl | |
0.6N HCl |
J402A | 1 × 15ml |
U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.
Resources
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