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Transcreener cGAMP cGAS分析法直接測量由環(huán)狀GMP-AMP合酶（cGAS，C6orf150、2'3'-cGAMP合酶）產(chǎn)生的環(huán)狀GMP-AMP （cGAMP）。通過測量cGAMP的產(chǎn)量，研究人員可以有效地確定 cGAS酶的活性。該測定法提供了一個強大的工具，可以篩選整個化合物庫中的cGAS調(diào)節(jié)劑，以 幫助找到針對cGAS-STING途徑的新療法。
真核細胞胞質(zhì)溶膠中DNA的存在是感染或細胞損傷的指示，并且在I型干擾素（IFN）誘導的驅動下引發(fā)強烈的免疫反應。cGAS是細胞質(zhì)中雙鏈DNA的非常靈敏的傳感器。DNA的結合激活了該酶，從而產(chǎn)生了*的環(huán)狀二核苷酸第二信使cGAMP，它充當STING（干擾素基因刺激劑）受體的激動劑，從而導致I型干擾素應答。
Transcreener cGAMP FP分析-了解更多信息
Transcreener cGAS分析法采用易于HTS使用，混合閱讀的格式。運行您的酶反應，添加 Transcreener試劑，然后閱讀板。該測定與96、384和1536孔格式兼容。
當使用標準曲線將原始數(shù)據(jù)轉換為cGAMP時，該測定法顯示線性。cGAS被雙鏈DNA激活，并需要ATP和GTP底物來生產(chǎn)cGAMP。
Transcreener cGAS分析法旨在以高通量格式篩選化合物庫。還可以使用該測定法進行隨訪SAR，從而輕松確定抑制劑的效價。
該測定法準確地證明了cGAS酶的特征。在GTP和ATP均存在的情況下，雙鏈DNA激活了產(chǎn)生cGAMP的酶。一個45個堿基對的ISD（干擾素刺激性DNA）和一個60個堿基對的HSV（單純皰疹病毒）DNA均可激活該酶。該測定法對cGAMP具有很高的親和力，即使存在其他核苷酸也可以檢測。
該測定顯示24小時以上的信號穩(wěn)定性。這樣的印版穩(wěn)定性對于大型的高通量篩選至關重要，由于大量樣品需要大量時間才能完成。
標準曲線包括ATP，GTP，dsDNA和cGAMP的滴定。在每個時間點讀取相同的板。
有興趣推進您的程序，但不想內(nèi)部進行分析嗎？我們的科學家可以提供幫助！BellBrook的科學家將利用其豐富的生物化學和酶學專業(yè)知識與您合作，并加快您的cGAS計劃。
| Component | Notes |
|---|---|
| cGAMP Antibody | Antibody specific to the dinucleotide cGAMP. |
| Tracer | When displaced, changes in fluorescence can be read in an FP or TR-FRET depending on the assay format. |
| Stop & Detect Buffer B, 10X | A buffer that can be used to stop enzyme reactions for an endpoint readout. |
| ATP | ATP can be used for the enzyme reaction and standard curve. |
| GTP | GTP can be used for the enzyme reaction and standard curve. |
| cGAMP | cGAMP can be used to create a standard curve. |
| Interferon Stimulatory DNA | ISD is a 45-bp oligomer used to activate the cGAS enzyme. |
| Component | Notes |
|---|---|
| cGAS Enzyme | You can purchase active cGAS enzyme directly through BellBrook Labs. |
| Enzyme Buffer | We routinely use enzyme buffer containing 20 mM TRIS (pH 7.5), 5 mM MgCl2, 0.01% Brij-35, and 1% DMSO (test compound solvent). Use the buffer that is a best fit for your enzyme conditions |
| Assay Plates | You can purchase assay plates directly through BellBrook Labs.
FP - An entirely black plate with a non-binding surface is required. We suggest Corning 384-well plates, Cat. #4514. TR-FRET - An entirely white plate with a non-binding surface is required. We suggest Corning 384-well plates Cat. #4513 |
| Cat. No. | Product Description | Price |
|---|---|---|
| 3024-1K | Transcreener cGAMP cGAS FP Assay (1,000 Assay, 384 Well) | $798.00 |
| 3024-10K | Transcreener cGAMP cGAS FP Assay (10,000 Assay, 384 Well) | $4,615.00 |
| 3024-100K | Transcreener cGAMP cGAS FP Assay (10 X 10,000 Assay, 384 Well) | $23,075.00 |
| 3025-1K | Transcreener cGAMP cGAS TR-FRET Assay (1,000 Assay, 384 Well) | $875.00 |
| 3025-10K | Transcreener cGAMP cGAS TR-FRET Assay (10,000 Assay, 384 Well) | $4,989.00 |
| 3025-100K | Transcreener cGAMP cGAS TR-FRET Assay (10 X 10,000 Assay, 384 Well) | $24,950.00 |
* For custom or bulk orders (over 100 μg) please contact us (info@bellbrooklabs.com) for a quote.
| Cat. No. | Product Description | Price |
|---|---|---|
| 2227 | Recombinant cGAS Enzyme, Active, Human, Full-Length, 10 µg | $399.00 |
| 2228 | Recombinant cGAS Enzyme, Active, Human, Full-Length, 100 µg | $1,398.00 |
* Includes enough enzyme for respective wells when used at 1.75 μg/mL (30 nM)
| Cat. No. | Product Description | Price |
|---|---|---|
| 3024-1K-B | Includes Transcreener cGAS FP Assay 1K Kit and 20 µg Human cGAS Enzyme | $1,275.00 |
| 3024-10K-B | Includes Transcreener cGAS FP Assay 10K Kit and 200 µg Human cGAS Enzyme | $6,670.00 |
| 3025-1K-B | Includes Transcreener cGAS TR-FRET Assay 1K Kit and 20 µg Human cGAS Enzyme | $1,345.00 |
| 3025-10K-B | Includes Transcreener cGAS TR-FRET Assay 10K Kit and 200 µg Human cGAS Enzyme | $6,995.00 |
| Cat. No. | Product Description | Price |
|---|---|---|
| 2233 | Corning 96-Well Black Assay Plates (#3686), 3-Pack (200+ Assays) | $40.00 |
| 2229 | Corning 384-Well Black Assay Plates (#4514), 3-Pack (1,000+ Assays) | $45.00 |
| 2230 | Corning 384-Well Black Assay Plates (#4514), 30-Pack (10,000+ Assays) | $400.00 |
| Cat. No. | Product Description | Price |
|---|---|---|
| 2234 | Corning 96-Well White Assay Plates (#3642), 3-Pack (200+ Assays) | $40.00 |
| 2231 | Corning 384-Well White Assay Plates (#4513), 3-Pack (1,000+ Assays) | $50.00 |
| 2232 | Corning 384-Well White Assay Plates (#4513), 30-Pack (10,000+ Assays) | $450.00 |
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