現(xiàn)貨產(chǎn)品磷酸化乙酰輔酶A(PaCoA)試劑盒齊一生物科技(上海)有限公司專業(yè)經(jīng)營ELISA試劑盒產(chǎn)品種類齊全、質(zhì)量可靠、*、靈敏度高、效果穩(wěn)定、易保存、操作簡單.到貨快、確保每一個ELISA試劑盒質(zhì)量.凡是購買我公司ELISA試劑盒免費代測、提供技術(shù)服務(wù)、5個工作日出實驗結(jié)果.歡迎新老客戶前來:
現(xiàn)貨產(chǎn)品磷酸化乙酰輔酶A(PaCoA)試劑盒ELISA試劑盒操作步驟
ELISA試劑盒使用前,將所有試劑充分混勻.不要使液體產(chǎn)生大量的泡沫,以免加樣時加入大量的氣泡,產(chǎn)生加樣上的誤差.
根據(jù)待測樣品數(shù)量加上標(biāo)準(zhǔn)品的數(shù)量決定所需的板條數(shù).每個標(biāo)準(zhǔn)品和空白孔建議做復(fù)孔.每個樣品根據(jù)自己的數(shù)量來定,能使用復(fù)孔的盡量做復(fù)孔.標(biāo)本用標(biāo)本稀釋液1:1稀釋后加入50ul于反應(yīng)孔內(nèi).
加入稀釋好后的標(biāo)準(zhǔn)品50ul于反應(yīng)孔、加入待測樣品50ul于反應(yīng)孔內(nèi).立即加入50ul的生物素標(biāo)記的抗體.蓋上膜板,輕輕振蕩混勻,37℃溫育1小時.
甩去孔內(nèi)液體,每孔加滿洗滌液,振蕩30秒,甩去洗滌液,用吸水紙拍干.重復(fù)此操作3次.如果用洗板機洗滌,洗滌次數(shù)增加一次.
每孔加入80ul的親和鏈酶素-HRP,輕輕振蕩混勻,37℃溫育30分鐘.
甩去孔內(nèi)液體,每孔加滿洗滌液,振蕩30秒,甩去洗滌液,用吸水紙拍干.重復(fù)此操作3次.如果用洗板機洗滌,洗滌次數(shù)增加一次.
每孔加入底物A、B各50ul,輕輕振蕩混勻,37℃溫育10分鐘.避免光照.
取出酶標(biāo)板,迅速加入50ul終止液,加入終止液后應(yīng)立即測定結(jié)果.
在450nm波長處測定各孔的OD值.
ELISA操作步驟
酶聯(lián)免疫吸附實驗材料,試劑,溶液
1.酶標(biāo)板,100μltip頭,1mlEp管,濕盒
2.包被稀釋液(0.05mol/L碳酸鈉-碳酸氫鈉buffer,pH9.6)
碳酸鈉0.15g,碳酸氫鈉0.29g,疊氮鈉0.02g,加雙蒸水100ml,調(diào)pH9.6
3.封閉液(5%小牛血清/PBS溶液)
小牛血清5ml,1*PBS(pH7.4)95ml
4.洗滌液(PBST,pH7.4)
NaCl0.8g,KH2PO40.02g,Na2HPO4.12H2O0.29g,KCl0.02g,Tween200.05ml,疊氮鈉0.01g,加雙蒸水100ml,調(diào)pH7.4
5.樣本稀釋液(PBS,pH7.4)
NaCl0.8g,KH2PO40.02g,Na2HPO4.12H2O0.29g,KCl0.02g,疊氮鈉0.01g,加雙蒸水100ml,調(diào)pH7.4
6.酶標(biāo)第2抗體(羊抗兔)稀釋范圍1:5,000-1:100,000
7.底物液(TMB-過氧化氫尿素溶液)
①底物液A:TMB20mg,無水乙醇10ml,加雙蒸水100ml.
②底物液B(0.1mol/L檸檬酸-0.2mol/L磷酸二氫鈉緩沖液,pH5.0-5.4)
Na2HPO41.46g,檸檬酸0.933g,0.75%過氧化氫尿素0.64ml,加三蒸水100ml,調(diào)pH5.0-5.4
③底物A和B按1:1混合即成TMB-過氧化氫尿素溶液.
8.終止液(2mol/LH2SO4溶液)雙蒸水200ml,濃硫酸34ml,(緩慢滴加并不斷攪拌),加雙蒸水300ml
9.0.9%生理鹽水
Anti-CCP (cyclic citrullinated peptide)
Anti-CCL5/RANTES(regulated on activation normal T cell expressed and secreted)
Anti-CCL20/MIP3αMacrophage inflammatory ptotein 3 alpha)
Anti-CCL11(Eotaxin 1)
Anti-CCL22/MDC(CC chemokine ABCD-1)
Anti-CCL21/6Ckine(Chemokine (C-C motif) ligand 21)
Anti-CCL22/MDC(small inducible cytokine A22 precursor)
Anti-CCL26(Eotaxin 3)
Anti-CCR-1(CC chemokine receptor 1)
Anti-CCR-2/5(CC chemokine receptor 2/5)
Anti-CCR-2A(C-C Chemokine receptor 2A)
Anti-CCR-2B(C-C Chemokine receptor 2B)
Anti-CCR-3(CC chemokine receptor 3)
Anti-CCR-4(C-C chemokine receptor 4)
Anti-CCR-6/CD196(C-C chemokine receptor type 6)
Anti-CXC-R6(CXC-chemokine receptor 6)
Anti-CCR-7(C-C chemokine receptor 7)
anti-CXCL10/IP-10
Anti-CCR-8(C-C chemokine receptor 8)
Anti-CCR-9(C-C chemokine receptor 9)
Anti-CCL28(CC Chemokine Ligand 28)
Anti-CCR-10(C-C chemokine receptor 10)
Anti-CCRK(Cell cycle related kinase)
Anti-CD10/CALLA (Neutral Endopeptidase)
Anti-CD105/Endoglin/TGF-β receptor
Anti-CD117/c-kit/SCFR
Anti-phospho-c-kit/SCFR/CD117(pTyr936)
Anti-CD133 antigen
Anti-CD138/Syndecan-1
Anti-CD146/MCAM
Anti-CD141(Thrombomodulin)
Anti-CD13/ANPEN(Aminopeptidase N)
Anti-CD14(cluster of differentiation antigen 14)
Anti-CD28
Anti-CD144/VE-cadherin/VECD(Vascularendothelial cell cadherin/VE-cadherin)VE-cadherin
Anti-CD147/EMMPRIN(Extracellular matrix metalloproteinase inducer)
Anti-CD166/ALCAM(activated leukocyte celladhesion molecule)
Anti-CD158a/KIR2DL1(Killer cell immunoglobulin-like receptor 2DL1)
Anti-CD158e/KIR3DL1(Killer cell immunoglobulin-like receptor 3DL1)
Anti-CD167a/DDR1
Anti-CD169/Siglec-1/sialoadhesin(Sn)(Sialic acid-binding Ig-like lectin 1)
Anti-CD177/NB1(Neutrophil Glycoprotein NB1)
Anti-CD17
Anti-CD18/LFA-1(Leukocyte Function Associated Anti-gen)
Anti-CD184/CXC-R4(chemokine receptor type 4)
Anti-CD19 (B-lymphocyte Anti-gen CD19 precursor)
Anti-CD20
Anti-CD22/fpc1/blcam(B lymphocyte cell adhesion molecule)
Anti-CD24
Anti-CD25/IL-2RA(Imterleukm-2 Receptor alpha)
Anti-CD122/IL-2RB(Imterleukm-2 Receptor beta)
Anti-CD272/BTLA(B and T lymphocyte attenuator)
Anti-CD2AP (CD2-associated protein)
Anti-CD3
Anti-CD31(PECAM-1)
Anti-CD31(PECAM-1)
Anti-CD33(human)
Anti-CD33/Siglec-3(mouse ret)
Anti-CD34
Anti-CD34(C-term)
Anti-CD38(mouse, rat)
Anti-CD38(to human)
Anti-CD36
Anti-CD4(mouse monoclonal)
Anti-CD4
Anti-CD7
Anti-CD8
Anti-CD40L(CD40 Ligand/TNFSF5/CD154)
Anti-CD44
Anti-CD45(LCA/gp180)
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