產品名稱:組(zu)織及(ji)血液(ye)酸性磷酸酶(ACP)活性檢測試劑(ji)盒
產品型號:BC2135
產品特點:組織及血液酸性磷酸酶(ACP)活性檢測試劑盒測定意義:ACP在酸性條件下催化磷酸單酯水解稱無機磷酸,常見于巨噬細胞的溶酶體內。ACP常用于前列腺癌的輔助診斷。測定原理:在酸性環境中,ACP催化磷酸苯二鈉水解生成苯酚,苯酚與4-氨基安替和鐵反應生成紅色亞醌衍生物,在510nm有特征光吸收;通過測定510nm吸光度增加速率,來計算ACP活性。
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組織及血液酸性磷酸酶(ACP)活性檢測試劑盒
ACP 在(zai)酸(suan)(suan)性(xing)(xing)條件下催化磷(lin)酸(suan)(suan)單酯(zhi)水解(jie)稱無機磷(lin)酸(suan)(suan),常(chang)見于巨噬(shi)細胞的(de)溶酶體內(nei)。ACP 常(chang)用于前(qian)列(lie)腺癌的(de)輔助診斷。在(zai)酸(suan)(suan)性(xing)(xing)環境中(zhong),ACP 催化磷(lin)酸(suan)(suan)苯二鈉水解(jie)生成苯酚,苯酚與 4-氨基安替和鐵(tie)反(fan)應生成紅色亞醌衍(yan)生物,在(zai) 510nm 有特征光吸收(shou);通過測定 510nm 吸光度增加(jia)速率(lv),來計算(suan) ACP 活性(xing)(xing)。
組織及血液酸性磷酸酶(ACP)活性檢測試劑盒
一、粗酶液提取: 稱取約 0.1g 組(zu)織,加提取液 1mL 充分研磨,4℃、10000rpm 離心 10min,取上清(qing)液待(dai)測。血液可直接用于測定(ding),如(ru)果(guo)濃度高的話,用提取液稀釋。
二、測定步驟:
1.分光光度計(ji)預(yu)熱 30 min 以上(shang),調節波長到(dao) 510 nm,蒸餾水調零。
2. 試(shi)(shi)劑(ji)一置于 37℃水浴中預熱 30 min 以上(shang)。試(shi)(shi)劑(ji)名稱(μL) 測(ce)定(ding)(ding)管(guan)(guan)(guan) 對照管(guan)(guan)(guan) 空白管(guan)(guan)(guan) 標準(zhun)管(guan)(guan)(guan)蒸餾水 - - 20 - 1μmol/mL 標準(zhun)品(pin) - - - 20 上(shang)清(qing)液 20 - - - 試(shi)(shi)劑(ji)一 40 40 40 40 試(shi)(shi)劑(ji)二 40 40 40 40 混勻后置于 37℃水浴中保(bao)溫 15min 試(shi)(shi)劑(ji)三 120 120 120 120 上(shang)清(qing)液 - 20 - - 混勻后于 510 nm 測(ce)定(ding)(ding)吸光度,分別記為 A 測(ce)定(ding)(ding)管(guan)(guan)(guan)、A 對照管(guan)(guan)(guan)、A 空白管(guan)(guan)(guan)、A 標準(zhun)管(guan)(guan)(guan)。
三(san)、ACP 活(huo)性(xing)(xing)計算(suan): 1.按蛋(dan)白(bai)濃(nong)度計算(suan)活(huo)性(xing)(xing)單位(wei)定(ding)(ding)義:37℃中(zhong)每毫克蛋(dan)白(bai)每分(fen)鐘催化產生(sheng) 1 μmol 酚。 ACP(U/mg pr)= [C 標(biao)(biao)(biao)(biao)準(zhun)品(pin)×(A 測(ce)定(ding)(ding)管(guan)(guan)(guan)(guan)(guan)(guan)-A 對(dui)(dui)照管(guan)(guan)(guan)(guan)(guan)(guan))÷(A 標(biao)(biao)(biao)(biao)準(zhun)管(guan)(guan)(guan)(guan)(guan)(guan)-A 空(kong)(kong)白(bai)管(guan)(guan)(guan)(guan)(guan)(guan))×V 樣(yang)(yang)(yang)]÷(Cpr×V 樣(yang)(yang)(yang))÷T = (A 測(ce)定(ding)(ding)管(guan)(guan)(guan)(guan)(guan)(guan)-A 對(dui)(dui)照管(guan)(guan)(guan)(guan)(guan)(guan))÷(A 標(biao)(biao)(biao)(biao)準(zhun)管(guan)(guan)(guan)(guan)(guan)(guan)-A 空(kong)(kong)白(bai)管(guan)(guan)(guan)(guan)(guan)(guan)) ÷Cpr 2.按樣(yang)(yang)(yang)本鮮(xian)重計算(suan)活(huo)性(xing)(xing)單位(wei)定(ding)(ding)義:37℃中(zhong)每克組(zu)織每分(fen)鐘催化產生(sheng) 1 μmol 酚。 ACP 活(huo)力(li)(U/g)= [C 標(biao)(biao)(biao)(biao)準(zhun)品(pin)×(A 測(ce)定(ding)(ding)管(guan)(guan)(guan)(guan)(guan)(guan)-A 對(dui)(dui)照管(guan)(guan)(guan)(guan)(guan)(guan))÷(A 標(biao)(biao)(biao)(biao)準(zhun)管(guan)(guan)(guan)(guan)(guan)(guan)-A 空(kong)(kong)白(bai)管(guan)(guan)(guan)(guan)(guan)(guan))×V 樣(yang)(yang)(yang)] ÷(W÷V 提取(qu)×V 樣(yang)(yang)(yang))÷T =0.067×(A 測(ce)定(ding)(ding)管(guan)(guan)(guan)(guan)(guan)(guan)-A 對(dui)(dui)照管(guan)(guan)(guan)(guan)(guan)(guan))÷(A 標(biao)(biao)(biao)(biao)準(zhun)管(guan)(guan)(guan)(guan)(guan)(guan)-A 空(kong)(kong)白(bai)管(guan)(guan)(guan)(guan)(guan)(guan))÷W 3.血(xue)(xue)液(ye)中(zhong) ACP 活(huo)性(xing)(xing)計算(suan)活(huo)性(xing)(xing)單位(wei)定(ding)(ding)義:37℃中(zhong)每毫升血(xue)(xue)液(ye)每分(fen)鐘催化產生(sheng) 1μmol 酚。 ACP 活(huo)力(li)(U)= [C 標(biao)(biao)(biao)(biao)準(zhun)品(pin)×(A 測(ce)定(ding)(ding)管(guan)(guan)(guan)(guan)(guan)(guan)-A 對(dui)(dui)照管(guan)(guan)(guan)(guan)(guan)(guan))÷(A 標(biao)(biao)(biao)(biao)準(zhun)管(guan)(guan)(guan)(guan)(guan)(guan)-A 空(kong)(kong)白(bai)管(guan)(guan)(guan)(guan)(guan)(guan))×V 樣(yang)(yang)(yang)] ÷V 樣(yang)(yang)(yang)÷T =0.067×(A 測(ce)定(ding)(ding)管(guan)(guan)(guan)(guan)(guan)(guan)-A 對(dui)(dui)照管(guan)(guan)(guan)(guan)(guan)(guan))÷(A 標(biao)(biao)(biao)(biao)準(zhun)管(guan)(guan)(guan)(guan)(guan)(guan)-A 空(kong)(kong)白(bai)管(guan)(guan)(guan)(guan)(guan)(guan)) C 標(biao)(biao)(biao)(biao)準(zhun)品(pin):1μmol/mL; V 樣(yang)(yang)(yang):加(jia)入(ru)反應體系中(zhong)上清液(ye)體積(ji)(mL),0.02mL;V 樣(yang)(yang)(yang)總:1mL; T:反應時間(min),15 min;V 提取(qu):加(jia)入(ru)提取(qu)液(ye)體積(ji),1mL;W:樣(yang)(yang)(yang)本鮮(xian)重,g。
組織及血液酸性磷酸酶(ACP)活性檢測試劑盒
提取(qu)液(ye):液(ye)體(ti) 50mL×1 瓶(ping),4℃保(bao)(bao)存(cun)(cun)。試(shi)(shi)劑一:液(ye)體(ti) 10mL×1 瓶(ping),4℃避光保(bao)(bao)存(cun)(cun)。試(shi)(shi)劑二(er):液(ye)體(ti) 10mL×1 瓶(ping),4℃避光保(bao)(bao)存(cun)(cun)。試(shi)(shi)劑三:液(ye)體(ti) 30mL×1 瓶(ping),4℃避光保(bao)(bao)存(cun)(cun),變成藍綠色不能使用。標準品(pin):液(ye)體(ti) 1mL×1 支,1μmol/mL 酚標準液(ye),4℃保(bao)(bao)存(cun)(cun)
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