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Transcreener cGAMP cGAS分析法直接測(cè)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(guò)測(cè)量cGAMP的產(chǎn)量，研究人員可以有效地確定 cGAS酶的活性。該測(cè)定法提供了一個(gè)強(qiáng)大的工具，可以篩選整個(gè)化合物庫(kù)中的cGAS調(diào)節(jié)劑，以 幫助找到針對(duì)cGAS-STING途徑的新療法。
真核細(xì)胞胞質(zhì)溶膠中DNA的存在是感染或細(xì)胞損傷的指示，并且在I型干擾素（IFN）誘導(dǎo)的驅(qū)動(dòng)下引發(fā)強(qiáng)烈的免疫反應(yīng)。cGAS是細(xì)胞質(zhì)中雙鏈DNA的非常靈敏的傳感器。DNA的結(jié)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(dāng)STING（干擾素基因刺激劑）受體的激動(dòng)劑，從而導(dǎo)致I型干擾素應(yīng)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(yùn)行您的酶反應(yīng)，添加 Transcreener試劑，然后閱讀板。該測(cè)定與96、384和1536孔格式兼容。
當(dāng)使用標(biāo)準(zhǔn)曲線將原始數(shù)據(jù)轉(zhuǎn)換為cGAMP時(shí)，該測(cè)定法顯示線性。cGAS被雙鏈DNA激活，并需要ATP和GTP底物來(lái)生產(chǎn)cGAMP。
Transcreener cGAS分析法旨在以高通量格式篩選化合物庫(kù)。還可以使用該測(cè)定法進(jìn)行隨訪SAR，從而輕松確定抑制劑的效價(jià)。
該測(cè)定法準(zhǔn)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(gè)45個(gè)堿基對(duì)的ISD（干擾素刺激性DNA）和一個(gè)60個(gè)堿基對(duì)的HSV（單純皰疹病毒）DNA均可激活該酶。該測(cè)定法對(duì)cGAMP具有很高的親和力，即使存在其他核苷酸也可以檢測(cè)。
該測(cè)定顯示24小時(shí)以上的信號(hào)穩(wěn)定性。這樣的印版穩(wěn)定性對(duì)于大型的高通量篩選至關(guān)重要，由于大量樣品需要大量時(shí)間才能完成。
標(biāo)準(zhǔn)曲線包括ATP，GTP，dsDNA和cGAMP的滴定。在每個(gè)時(shí)間點(diǎn)讀取相同的板。
有興趣推進(jìn)您的程序，但不想內(nèi)部進(jìn)行分析嗎？我們的科學(xué)家可以提供幫助！BellBrook的科學(xué)家將利用其豐富的生物化學(xué)和酶學(xué)專業(yè)知識(shí)與您合作，并加快您的cGAS計(jì)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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