 |
 |
 |
 |
 |
 |
||
 |
 |
 |
 |
 |
| 1 | 1/1 | 返回列表 |
| 查看: 2144 | 回復(fù): 0 | ||
laoyongmin新蟲 (初入文壇)
|
[求助]
怎么測表觀Km常數(shù)？
|
|
目前在純化His-tagged蛋白，馬上做酶性質(zhì)分析，想測表觀Km（apparent kinetic constants），參考文獻(xiàn)（和我一樣的酶）：“For determination of apparent kinetic constants, eight concentrations each of the substrates DMADP (0–100 uM), GDP (0–50 uM), FDP (0–50 uM) or GGDP (0–50 uM) were used in the presence of a fixed IDP concentration (100 uM). The assay mixtures contained 25 mM MOPSO (pH 7.2), 10 mM MgCl2 and 10% v/v glycerol, plus IDP and DMADP, GDP, FDP or GGDP and 1 ug purified AtIDS9 per assay in a 200 uL volume. The reaction mixture was incubated in glass vials at 20°C for times ranging from 0–2 min in 10 sec intervals with shaking using an Eppendorf Thermomixer Comfort (www.eppendorf.com), and the reaction was stopped by immediately freezing the sample in liquid nitrogen. To determine the GFDP concentration, samples were analyzed via UPLC/MS using a calibration curve”，這里寫的8個(gè)濃度和反應(yīng)時(shí)間（0-2 min）該如何設(shè)置？文獻(xiàn)的結(jié)果如圖所示，有些只有7和點(diǎn)，不是有8個(gè)濃度嗎？圖中的1/V和1/S是怎樣算出來的？ Figure S2: Lineweaver-Burk plots for calculation of apparent in vitro Km values of AtIDS9 with different substrates. The double reciprocal velocity (1/v) and substrate concentrations (1/[S]) are plotted to determine the apparent Km values. A. DMADP (0-100 µM) + 100 µM IDP used as substrate B. GDP (0-50 µM) + 100 µM IDP used as substrate C. FDP (0-50 µM) + 100 µM IDP used as substrate. D. GGDP (0-15 µM) + 100 µM IDP used as substrate. E. IDP (0-40 µM) + 50 µM GGDP used as substrate. 11.jpg |
回復(fù)此樓» 猜你喜歡
085600材料與化工
已經(jīng)有6人回復(fù)
-
281求調(diào)劑（0805）
已經(jīng)有24人回復(fù)
-
265求調(diào)劑
已經(jīng)有8人回復(fù)
-
0817 化學(xué)工程 299分求調(diào)劑 有科研經(jīng)歷 有二區(qū)文章
已經(jīng)有21人回復(fù)
-
295材料求調(diào)劑，一志愿武漢理工085601專碩
已經(jīng)有4人回復(fù)
-
0856調(diào)劑，是學(xué)校就去
已經(jīng)有9人回復(fù)
-
298-一志愿中國農(nóng)業(yè)大學(xué)-求調(diào)劑
已經(jīng)有9人回復(fù)
-
求調(diào)劑
已經(jīng)有4人回復(fù)
-
學(xué)校已經(jīng)提交到NSFC，還能修改嗎？
已經(jīng)有7人回復(fù)
-
材料學(xué)碩297已過四六級求調(diào)劑推薦
已經(jīng)有6人回復(fù)
相關(guān)版塊跳轉(zhuǎn)
我要訂閱樓主 laoyongmin 的主題更新
| 1 | 1/1 | 返回列表 |
| 最具人氣熱帖推薦 [查看全部] | 作者 | 回/看 | 最后發(fā)表 | |
|---|---|---|---|---|
|
[考研] 281求調(diào)劑（0805） +13 | 煙汐憶海 2026-03-16 | 24/1200 |
|
|---|---|---|---|---|
|
[考研] 0856調(diào)劑，是學(xué)校就去 +8 | sllhht 2026-03-19 | 9/450 |
|
|
|
[考研] 一志愿武漢理工材料工程專碩調(diào)劑 +7 | Doleres 2026-03-19 | 7/350 |
|
|
|
[考研] 085410人工智能專碩317求調(diào)劑（0854都可以） +4 | xbxudjdn 2026-03-18 | 4/200 |
|
|
|
[考研] 288求調(diào)劑 +15 | 于海海海海 2026-03-19 | 15/750 |
|
|
|
[考研] 085601材料工程專碩求調(diào)劑 +10 | 慕寒mio 2026-03-16 | 10/500 |
|
|
|
[考研] 0817調(diào)劑 +3 | 沒有答案_ 2026-03-14 | 3/150 |
|
|
|
[考研] 本科鄭州大學(xué)物理學(xué)院，一志愿華科070200學(xué)碩，346求調(diào)劑 +4 | 我不是一根蔥 2026-03-18 | 4/200 |
|
|
|
[考研] 一志愿華中科技大學(xué)，080502，354分求調(diào)劑 +4 | 守候夕陽CF 2026-03-18 | 4/200 |
|
|
|
[考研] 311求調(diào)劑 +11 | 冬十三 2026-03-15 | 12/600 |
|
|
|
[考研] 收復(fù)試調(diào)劑生 +4 | 雨后秋荷 2026-03-18 | 4/200 |
|
|
|
[考研]
 0854，計(jì)算機(jī)類招收調(diào)劑
+3
|
胡辣湯放糖 2026-03-15 | 6/300 |
|
|
|
[考研] 環(huán)境工程調(diào)劑 +8 | 大可digkids 2026-03-16 | 8/400 |
|
|
|
[考博] 26博士申請 +3 | 1042136743 2026-03-17 | 3/150 |
|
|
|
[考研] 材料專碩326求調(diào)劑 +6 | 墨煜姒莘 2026-03-15 | 7/350 |
|
|
|
[考研] 東南大學(xué)364求調(diào)劑 +5 | JasonYuiui 2026-03-15 | 5/250 |
|
|
|
[考研] 318求調(diào)劑 +3 | Yanyali 2026-03-15 | 3/150 |
|
|
|
[考研] 070300化學(xué)學(xué)碩求調(diào)劑 +6 | 太想進(jìn)步了0608 2026-03-16 | 6/300 |
|
|
|
[考研] 070303 總分349求調(diào)劑 +3 | LJY9966 2026-03-15 | 5/250 |
|
|
|
[考研] 22408總分284求調(diào)劑 +3 | InAspic 2026-03-13 | 3/150 |
|