Name: 괴사, 세포사멸 및 혈소판 활성화의 특징을 보이는 Procoagulant Platelets의 포괄적인 분석 (동영상) | JoVE Journal
Uploaded: 2025-05-23T14:00:00.000+00:00
Duration: 4 min 37 s
Description: 학습하세요 혈소판 대사와 활성화 상태 분석을 통해 혈전증 바이오마커를 식별하는 방법을. 이 프로토콜은 procoagulant platelets의 기능적 특성을 규명하여 치료 표적을 탐색하는 데 활용됩니다.

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5 minutes

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<li>Kulkarni, P. P., Sonkar, V. K., Gautam, D., Dash, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((AMPK+inhibition+protects+against+arterial+thrombosis+while+sparing+hemostasis+through+differential+modulation+of+platelet+responses%5BTitle%5D)+AND+%22Thromb+Res%22%5BJournal%5D)">AMPK inhibition protects against arterial thrombosis while sparing hemostasis through differential modulation of platelet responses.</a> Thromb Res. 196, 175-185 (2020).</li>
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<li>Kholmukhamedov, A., Janecke, R., Choo, H. -J., Jobe, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+mitochondrial+calcium+uniporter+regulates+procoagulant+platelet+formation%5BTitle%5D)+AND+%22J+Thromb+Haemost%22%5BJournal%5D)">The mitochondrial calcium uniporter regulates procoagulant platelet formation.</a> J Thromb Haemost. 16 (11), 2315-2321 (2018).</li>
<li>Choo, H. -J., Saafir, T. B., Mkumba, L., Wagner, M. B., Jobe, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+calcium+and+reactive+oxygen+species+regulate+agonist-initiated+platelet+phosphatidylserine+exposure%5BTitle%5D)+AND+%22Arterioscler+Thromb+Vasc+Biol%22%5BJournal%5D)">Mitochondrial calcium and reactive oxygen species regulate agonist-initiated platelet phosphatidylserine exposure.</a> Arterioscler Thromb Vasc Biol. 32 (12), 2946-2955 (2012).</li>
<li>Jobe, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Critical+role+for+the+mitochondrial+permeability+transition+pore+and+cyclophilin+D+in+platelet+activation+and+thrombosis%5BTitle%5D)+AND+%22Blood%22%5BJournal%5D)">Critical role for the mitochondrial permeability transition pore and cyclophilin D in platelet activation and thrombosis.</a> Blood. 111 (3), 1257-1265 (2008).</li>
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<li>Lopez, J. J., Salido, G. M., Pariente, J. A., Rosado, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Thrombin+induces+activation+and+translocation+of+Bid%2C+Bax+and+Bak+to+the+mitochondria+in+human+platelets%5BTitle%5D)+AND+%22J+Thromb+Haemost%22%5BJournal%5D)">Thrombin induces activation and translocation of Bid, Bax and Bak to the mitochondria in human platelets.</a> J Thromb Haemost. 6 (10), 1780-1788 (2008).</li>
<li>Wolf, B. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Calpain+functions+in+a+caspase-independent+manner+to+promote+apoptosis-like+events+during+platelet+activation%5BTitle%5D)+AND+%22Blood%22%5BJournal%5D)">Calpain functions in a caspase-independent manner to promote apoptosis-like events during platelet activation.</a> Blood. 94 (5), 1683-1692 (1999).</li>
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<li>Schoenwaelder, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Two+distinct+pathways+regulate+platelet+phosphatidylserine+exposure+and+procoagulant+function%5BTitle%5D)+AND+%22Blood%22%5BJournal%5D)">Two distinct pathways regulate platelet phosphatidylserine exposure and procoagulant function.</a> Blood. 114 (3), 663-666 (2009).</li>
<li>Abbasian, N., Millington-Burgess, S. L., Chabra, S., Malcor, J. -D., Harper, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Supramaximal+calcium+signaling+triggers+procoagulant+platelet+formation%5BTitle%5D)+AND+%22Blood+Adv%22%5BJournal%5D)">Supramaximal calcium signaling triggers procoagulant platelet formation.</a> Blood Adv. 4 (1), 154-164 (2020).</li>
<li>Le, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+inner+activation+gate+controls+TMEM16F+phospholipid+scrambling%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">An inner activation gate controls TMEM16F phospholipid scrambling.</a> Nat Commun. 10 (1), 1846(2019).</li>
<li>Yang, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((TMEM16F+forms+a+Ca2%2B-activated+cation+channel+required+for+lipid+scrambling+in+platelets+during+blood+coagulation%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">TMEM16F forms a Ca2+-activated cation channel required for lipid scrambling in platelets during blood coagulation.</a> Cell. 151 (1), 111-122 (2012).</li>
<li>Mattheij, N. J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dual+mechanism+of+integrin+closure+in+procoagulant+platelets%5BTitle%5D)+AND+%22J+Biol+Chem%22%5BJournal%5D)">Dual mechanism of integrin closure in procoagulant platelets.</a> J Biol Chem. 288 (19), 13325-13336 (2013).</li>
<li>Liu, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrially+mediated+integrin+%CE%B1iIb%CE%B23+protein+inactivation+limits+thrombus+growth%5BTitle%5D)+AND+%22J+Biol+Chem%22%5BJournal%5D)">Mitochondrially mediated integrin αiIbβ3 protein inactivation limits thrombus growth.</a> J Biol Chem. 288 (42), 30672-30681 (2013).</li>
<li>van Kruchten, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Both+TMEM16F-dependent+and+TMEM16F-independent+pathways+contribute+to+phosphatidylserine+exposure+in+platelet+apoptosis+and+platelet+activation%5BTitle%5D)+AND+%22Blood%22%5BJournal%5D)">Both TMEM16F-dependent and TMEM16F-independent pathways contribute to phosphatidylserine exposure in platelet apoptosis and platelet activation.</a> Blood. 121 (10), 1850-1857 (2013).</li>
<li>Goelz, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Platelets+express+adaptor+proteins+of+the+extrinsic+apoptosis+pathway+and+can+activate+caspase-8%5BTitle%5D)+AND+%22PLOS+ONE%22%5BJournal%5D)">Platelets express adaptor proteins of the extrinsic apoptosis pathway and can activate caspase-8.</a> PLOS ONE. 16 (1), e0244848(2021).</li>
<li>Shlomovitz, I., Speir, M., Gerlic, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Flipping+the+dogma+-+phosphatidylserine+in+non-apoptotic+cell+death%5BTitle%5D)+AND+%22Cell+Commun+Signal%22%5BJournal%5D)">Flipping the dogma - phosphatidylserine in non-apoptotic cell death.</a> Cell Commun Signal. 17 (1), 139(2019).</li>
<li>Nadine, J. A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Coated+platelets+function+in+platelet-dependent+fibrin+formation+via+integrin+%CE%B1IIb%CE%B23+and+transglutaminase+factor+XIII%5BTitle%5D)+AND+%22Haematologica%22%5BJournal%5D)">Coated platelets function in platelet-dependent fibrin formation via integrin αIIbβ3 and transglutaminase factor XIII.</a> Haematologica. 101 (4), 427-436 (2016).</li>
<li>Michelson, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Platelet+activation+by+thrombin+can+be+directly+measured+in+whole+blood+through+the+use+of+the+peptide+GPRP+and+flow+cytometry%3A+Methods+and+clinical+applications%5BTitle%5D)+AND+%22Blood+Coagul+Fibrinolysis%22%5BJournal%5D)">Platelet activation by thrombin can be directly measured in whole blood through the use of the peptide GPRP and flow cytometry: Methods and clinical applications.</a> Blood Coagul Fibrinolysis. 5 (1), 121-131 (1994).</li>
<li>Taylor, M. R., Couto, J. R., Scallan, C. D., Ceriani, R. L., Peterson, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Lactadherin+%28formerly+BA46%29%2C+a+membrane-associated+glycoprotein+expressed+in+human+milk+and+breast+carcinomas%2C+promotes+Arg-Gly-Asp+%28RGD%29-dependent+cell+adhesion%5BTitle%5D)+AND+%22DNA+Cell+Biol%22%5BJournal%5D)">Lactadherin (formerly BA46), a membrane-associated glycoprotein expressed in human milk and breast carcinomas, promotes Arg-Gly-Asp (RGD)-dependent cell adhesion.</a> DNA Cell Biol. 16 (7), 861-869 (1997).</li>
<li>Andersen, M. H., Graversen, H., Fedosov, S. N., Petersen, T. E., Rasmussen, J. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Functional+analyses+of+two+cellular+binding+domains+of+bovine+lactadherin%5BTitle%5D)+AND+%22Biochemistry%22%5BJournal%5D)">Functional analyses of two cellular binding domains of bovine lactadherin.</a> Biochemistry. 39 (20), 6200-6206 (2000).</li>
<li>Tan, C. W., Bourcy, M., Pasalic, L., Chen, V. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Flow+cytometry+assessment+of+procoagulant+platelets+using+a+dithiol-reactive+probe+functional+disulfide+bonds%3A+Methods+and+protocols%5BTitle%5D)+AND+%22Methods+Mol+Biol%22%5BJournal%5D)">Flow cytometry assessment of procoagulant platelets using a dithiol-reactive probe functional disulfide bonds: Methods and protocols.</a> Methods Mol Biol. 1967, 305-321 (2019).</li>
</ol>

<table><tbody><tr><td>산성 시트레이트 덱스트로오스 (ACD) 용액 (1000 mL용)</td><td></td><td></td><td>Tri- 나트륨 시트레이트- &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 22 g 구연산-&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 8 g 포도당-&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 24.5 g 물-&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 최대 용량은 1000 mL입니다&nbsp;</td></tr><tr><td>Alexa Fluor 488 단백질 라벨링 키트</td><td>Invitrogen</td><td>A10235</td><td></td></tr><tr><td>APC 마우스 Anti-Human CD62P</td><td>BD Pharmingen</td><td>550888</td><td></td></tr><tr><td>소 프로트롬빈</td><td>Prolytix</td><td>BCP-1010</td><td></td></tr><tr><td>버퍼 A (혈소판 준비)</td><td></td><td></td><td>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; M.W&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. 1X에서&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 100 mL 10X 용액 Hepes&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 238.30&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 20 미디엄&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 4.766 g NaCl&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 58.44&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 134 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 7.83 g KCl&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 74.55&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.9 밀리미터&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 216.19 mg MgCl2&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 203.30&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 203.30 mg NaH2PO4&nbsp; &nbsp; &nbsp; 156.01&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 0.34 제곱미터&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 53.04 mg NaHCO3&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 84 01&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 12 미디미터 &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; pH를 6.2 로 조정한 후 1.01 g 물을 100 mL로 추출하고, 혈소판 준비 직전에 Milli Q 물로 10X 용액을 1:10(v/v)로 희석하여 다음을 보충해야 하는 1X 용액을 얻습니다 &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. 1X에서&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 ml 1X 용액 EGTA &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 밀리미터; &nbsp; &nbsp; &nbsp; &nbsp; 10 추가 &mu; L (1:100 v/v) 100 mM EGTA&nbsp; 포도당 &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5 밀리미터; &nbsp; &nbsp; &nbsp; 5 추가 &mu; L (1:200 v/v) 1 M 포도당 PGE1 &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 3 &뮤; 미디엄 &nbsp; &nbsp; &nbsp; &nbsp; 3 추가 &mu; L (1:333 v/v) 1 mM PGE1 용액</td></tr><tr><td>버퍼 B (혈소판 제제)</td><td></td><td></td><td>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; M.W&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. 1X에서&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 100 mL 10X 용액 Hepes&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 238.30&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 20 미디엄&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 4.766 g NaCl&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 58.44&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 134 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 7.83 g KCl&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 74.55&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.9 밀리미터&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 216.19 mg MgCl2&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 203.30&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 203.30 mg NaH2PO4&nbsp; &nbsp; &nbsp; 156.01&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 0.34 제곱미터&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 53.04 mg NaHCO3&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 84 01&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 12 미디미터 &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; pH를 7.4 로 조정한 후 1.01 g 물을 100 mL에 넣고 혈소판 준비 직전에 Milli Q 물로 10X 용액을 1:10 (v/v)로 희석하여 다음을 보충해야 하는 1X 용액을 얻습니다 &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. 1X에서&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 ml 1X 용액 포도당 &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5 밀리미터; &nbsp; &nbsp; &nbsp; 5 추가 &mu; L (1:200 v/v) 1 M 포도당&nbsp;</td></tr><tr><td>CaCl2 (0.5 M) (50 ml의 경우)</td><td></td><td></td><td>CaCl2.2H2O의 분자량 = 147.02 50 ml Milli Q 물에 CaCl2.2H2O 3.675 g을 용해 CellEvent</td></tr><tr><td>Caspase-3/7 검출 시약 Green</td><td>Invitrogen</td><td>C10423</td><td>Apoptosis 검출 시약</td></tr><tr><td>Convulxin</td><td>Enzo Life Sciences</td><td>ALX-350-100</td><td></td></tr><tr><td>EDTA (0.5 M; pH 8) (100 mL 용)</td><td></td><td></td><td>EDTA Na2.2H2 O의 분자량 : 372.24g 18.612g의 무게를 측정하고 50ml Milli Q 물에 현탁시키고 pH (pH ~ 4) 를 확인하고 교반하면서 10N NaOH를 천천히 첨가하고 pH. 를 모니터링합니다. EDTA는 pH 7 부근에서 용해를 시작하고 pH 8. 에서 완전히 용해됩니다. Milli Q 물로 100ml로 부피를 구성합니다.</td></tr><tr><td>EGTA (100 mM; pH 7.4) (100 mL의 경우)</td><td></td><td></td> <td>분자량 : 380.4 3.804 ml Milli Q 물에 50 g EGTA를 넣고 pH (pH ~ 3) 확인 교반하면서 10 N NaOH를 적> 첨가 및 pH <7.0에서 < EGTA가 용해되는 pH 7.0 (대략) (약) br / >로 pH를 7.4 로 조정 Milli Q 물로</td></tr> <tr><td>100ml로 부피를 구성합니다.FITC Mouse Anti-Human PAC-1</td><td>BD</td><td>340507</td><td></td></tr><tr><td>FITC-IETD-FMK Caspase 8 (active) staining kit</td><td>Abcam</td><td>ab65614</td><td></td></tr><tr><td>Mitotracker Red CMXRos (mitochindria labeling dye)</td><td>Invitrogen</td><td>M7512</td><td>Stock= 1 mM (50 &micro; g는 90 &micro에 용해됩니다. l DMSO) 하위 재고= 100 &마이크로; M (10 &마이크로; l 재고 + 90 &micro; l DMSO) 작업 농도= 500nM (0.5 &마이크로; 100 &micro에 있는 l; l)</td></tr><tr><td>PE Annexin V</td><td>BD Pharmingen</td><td>560930</td><td></td></tr><tr><td>프로스타글란딘 E1</td><td>시그마</td><td>P5515</td><td>스톡= 20 mM (141 &micro에 용해된 1 mg; L DMSO) 하위 재고= 1 mM (10 &micro; l 재고 + 190 &micro; L DMSO) 작업 농도= 3 &마이크로; M (3 &마이크로; L in 1 mL)</td></tr><tr><td>Rhod-2 AM</td><td>Invitrogen</td><td>R1244</td><td>스톡= 5 mM (178 &micro에 용해된 1 mg; L DMSO) 하위 재고= 100 &마이크로; M (10 &마이크로; L 주식 + 90 &micro; L DMSO) 작업 농도= 500 nM (0.5 &마이크로; L 100 &마이크로; L)</td></tr><tr><td>인간 플라즈마</td><td>Sigma</td><td>T7572</td><td></td>의 트롬빈</tr></tbody></table>

괴사, 세포사멸 및 혈소판 활성화의 특징을 보이는 procoagulant platelets의 포괄적인 분석

Comprehensive Analysis of Procoagulant Platelets Exhibiting Features of Necrosis, Apoptosis and Platelet Activation

활성화된 혈소판의 일부는 포스파티딜세린(PS)과 P-셀렉틴의 표면 발현이 특징적으로 증가하면서 "응고제"로 변하는데, 이는 PS 노출에만 양성인 "자가사멸" 혈소판 및 P-셀렉틴 발현에 대해 양성인 "증식" 혈소판과 구별됩니다. 우리는 트롬빈이 FITC-anti-CD62P 항체 및 PE-Annexin V의 결합에 의해 검출된 P-selectin 및 PS 발현 모두에 대해 양성인 procoagulant 혈소판의 비율을 용량 의존적 증가를 유도한다는 것을 발견했습니다(그림 2). procoagulant platelets의 생성은 작용제에 의해 유도된 cytosolic calcium 상승에 따라 내부 미토콘드리아 막을 가로지르는 전기화학적 구배를 따라 미토콘드리아 칼슘 유입18,19 및 그에 따른 cyclophilin D (CypD) 의존성 미토콘드리아 투과성 전이 공극 (mP...

괴사, 세포사멸 및 혈소판 활성화의 특징을 보이는 Procoagulant Platelets의 포괄적인 분석

Platelets circulating in the bloodstream are relatively quiescent but become "activated" upon encountering stimulants or "agonists" at the site of blood ...

comprehensive-analysis-procoagulant-platelets-exhibiting-features

Research

연구

JoVE Journal

Medicine

의학

여기 혈관 염증 및 라이브 쥐 혈전 형성되는 동안 활성화 내피에 heterotypic 혈소판 호중구 상호 작용을 시각화하는 형광 intravital 현미경의 실험 기술을보고합니다. 이 현미경 기술은 혈관 질환의 분자 메커니즘을 연구 할 수 있고 pathophysiological 조건에 따라 pharmacologic 요원을 테스트 할 가치가 있습니다.

라이브 마우스의 혈관 염증과 혈전 형성하는 동안 활성화 내피에 Heterotypic 혈소판 - 호중구 상호 작용의 실시간 영상

이 연구는 혈소판의 부착, 퍼짐 및 유속 하의 분비를 연구하기위한 형광 현미경 기반의 방법을 설명합니다. 이 다목적 플랫폼은 혈전증과 지혈에 대한 기계 연구를 위해 혈소판 기능을 조사 할 수있게합니다.

흐름하에있는 혈소판 탈식 및 분비의 라이브 셀 이미징

프로응고제 혈소판 형성은 혈전증 위험 증가와 상관관계가 있습니다. 여기에는 인간 혈액에서 세척된 혈소판을 분리하기 위한 정확한 프로토콜이 제시되어 있으며, 이는 프로코아응고제 혈소판의 특징인 포스파티딜세린 및 미세소포 방출의 노출을 정량화하기 위한 것입니다.

인간 정제 혈소판에서 포스파티딜세린 노출 및 미세소포 방출을 측정하여 응고제 혈소판 특성 분석

혈류 하의 혈소판 기능을 평가하고 시뮬레이션된 지혈 소생술을 외상 및 수혈 의학에 응용하는 미세유체 장치를 사용하여 모델링할 수 있습니다.

혈소판 기능 평가에서의 미세유체역학(Microfluidics in Assessing Platelet Function)

혈소판에 대한 다색 유세포 분석은 고전적인 혈소판 하위 집단 내에서 휴지 혈소판, 응집성 혈소판, 단백질 응고제 및 자가사멸성 혈소판과 같은 새로운 혈소판 하위 유형을 식별할 수 있습니다. 이를 통해 다양한 혈소판 작용제에 의해 유도된 다양한 하위 집단 패턴을 비교할 수 있습니다. 여기에서는 multi-color 유세포 분석 패널을 설정하는 절차를 자세히 설명합니다.

Multi-color 유세포분석법을 통한 혈소판 하위 모집단 분석

이 연구는 전단 응력을 이용해 혈전 형성을 유도하고, 여러 차원의 판독을 통해 혈전을 특성화하는 마이크로유체 분석법을 설명합니다. 이 검사법은 혈액 샘플의 혈전형성 경향을 측정할 수 있어 질병 진단, 신약 개발, 혈전증과 관련된 기전적 연구에도 유용합니다.

혈전 프로파일링 분석법: 생체역학적 혈전 생성을 포괄적으로 특성화하기 위한 마이크로플루이딕스 기반 플랫폼

흉부 종양학 데이터베이스는 translational 연구의 목적을 위해 임상 실험 데이터에 대한 포괄적인 저장소 역할을하도록 개발되었습니다. 데이터베이스는 흉부 종양학 연구 프로그램의 translational 암 연구자를 제공합니다. 이 데이터베이스는 다른 암 모델뿐만 아니라 다른 사람의 질병에 적용할 수 있습니다.

종합 흉부 종양학 데이터베이스의 생성 - Translational 연구 도구

음성을 지원하는 생리적 메커니즘의 목적 평가는 ALS와 사람에서 질병 발병과 진행을 모니터하고 임상 실험에서 치료 효과를 계량이 필요합니다. 이 비디오에서는, 우리는 임상 인구의 음성 모터 성능을 quantifying에 대한 포괄적인, 계측 기반 프로토콜을 제시한다.

루게릭병에 안구 장애의 종합 평가 (ALS)에 대한 프로토콜

암 세포에 의해 내피 세포 단층의 침략을 측정하는 실시간 전기 임피던스 기반 기술

RNA 간섭 (RNAi)는 유전자 녹아웃 이상의 많은 장점을 보유하고 광범위하게 유전자 기능 연구의 도구로 사용되었습니다. DNA를 벡터 기반의 RNAi 기술의 발명은 장기적으로 가능한 inducible 유전자 최저했고, 또한 유전자 입을의 가능성을...

암 관련 유전자 기능을 연구하기 DNA를 벡터 기반의 RNA 간섭

학습하세요 혈소판 대사와 활성화 상태 분석을 통해 혈전증 바이오마커를 식별하는 방법을. 이 프로토콜은 procoagulant platelets의 기능적 특성을 규명하여 치료 표적을 탐색하는 데 활용됩니다.

괴사, 세포사멸 및 혈소판 활성화의 특징을 보이는 Procoagulant Platelets의 포괄적인 분석

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