FITC膜聯蛋白V細胞凋亡檢測試劑盒（BD Pharmingen）描述

凋(diao)(diao)亡(wang)(wang)(wang)(wang)(wang)是在(zai)胚胎(tai)發(fa)育過程(cheng)中發(fa)生的(de)(de)(de)正(zheng)常(chang)生理過程(cheng)以(yi)及組織穩態的(de)(de)(de)維持。凋(diao)(diao)亡(wang)(wang)(wang)(wang)(wang)程(cheng)序的(de)(de)(de)特(te)征是某(mou)些(xie)形態學特(te)征，包(bao)括(kuo)質(zhi)(zhi)膜(mo)(mo)不對(dui)稱(cheng)和(he)附(fu)著，細(xi)(xi)胞(bao)(bao)質(zhi)(zhi)和(he)細(xi)(xi)胞(bao)(bao)核(he)(he)的(de)(de)(de)縮合以(yi)及DNA的(de)(de)(de)核(he)(he)小(xiao)體間裂解。質(zhi)(zhi)膜(mo)(mo)的(de)(de)(de)損失是zui早(zao)的(de)(de)(de)特(te)征之一(yi)。在(zai)凋(diao)(diao)亡(wang)(wang)(wang)(wang)(wang)細(xi)(xi)胞(bao)(bao)中，膜(mo)(mo)磷(lin)(lin)脂磷(lin)(lin)脂酰(xian)絲氨酸（PS）從質(zhi)(zhi)膜(mo)(mo)的(de)(de)(de)內部和(he)外部葉片(pian)轉位，從而將PS暴露于外部細(xi)(xi)胞(bao)(bao)環(huan)境。膜(mo)(mo)聯(lian)蛋(dan)白(bai)(bai)(bai)V是一(yi)種35-36 kDa的(de)(de)(de)Ca2 +依(yi)賴性(xing)磷(lin)(lin)脂結合蛋(dan)白(bai)(bai)(bai)，對(dui)PS具(ju)有高親和(he)力，并與暴露的(de)(de)(de)PS結合細(xi)(xi)胞(bao)(bao)。膜(mo)(mo)聯(lian)蛋(dan)白(bai)(bai)(bai)V可(ke)以(yi)與包(bao)括(kuo)FITC的(de)(de)(de)熒光染料(liao)綴合。該格式保留了對(dui)PS的(de)(de)(de)高親和(he)力，因此用作對(dui)正(zheng)在(zai)進行細(xi)(xi)胞(bao)(bao)凋(diao)(diao)亡(wang)(wang)(wang)(wang)(wang)的(de)(de)(de)細(xi)(xi)胞(bao)(bao)的(de)(de)(de)流式細(xi)(xi)胞(bao)(bao)分析的(de)(de)(de)敏感探針。由于PS的(de)(de)(de)外化(hua)發(fa)生在(zai)凋(diao)(diao)亡(wang)(wang)(wang)(wang)(wang)的(de)(de)(de)早(zao)期階段(duan)，FITC膜(mo)(mo)聯(lian)蛋(dan)白(bai)(bai)(bai)V染色可(ke)以(yi)比基于核(he)(he)變(bian)化(hua)如DNA片(pian)段(duan)化(hua)的(de)(de)(de)測定在(zai)早(zao)期鑒(jian)定凋(diao)(diao)亡(wang)(wang)(wang)(wang)(wang)。

FITC膜(mo)(mo)(mo)(mo)(mo)聯蛋(dan)(dan)白(bai)(bai)V染(ran)色先于膜(mo)(mo)(mo)(mo)(mo)完(wan)整性(xing)(xing)(xing)(xing)(xing)的(de)(de)(de)(de)喪失(shi)，伴隨著由(you)凋(diao)亡(wang)(wang)(wang)(wang)(wang)或壞死(si)(si)過程引起的(de)(de)(de)(de)細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)死(si)(si)亡(wang)(wang)(wang)(wang)(wang)階(jie)段。因此，FITC膜(mo)(mo)(mo)(mo)(mo)聯蛋(dan)(dan)白(bai)(bai)V的(de)(de)(de)(de)染(ran)色通常與重要染(ran)料（如碘化丙啶（PI）或7-氨基放線菌素（7-AAD））結(jie)合(he)使(shi)用，以使(shi)研究(jiu)人(ren)員能夠鑒定(ding)早(zao)期(qi)(qi)凋(diao)亡(wang)(wang)(wang)(wang)(wang)細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)（PI陰性(xing)(xing)(xing)(xing)(xing)，FITC膜(mo)(mo)(mo)(mo)(mo)聯蛋(dan)(dan)白(bai)(bai)V正）。具有(you)完(wan)整膜(mo)(mo)(mo)(mo)(mo)的(de)(de)(de)(de)活細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)排除PI，使(shi)死(si)(si)亡(wang)(wang)(wang)(wang)(wang)和(he)受損細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)的(de)(de)(de)(de)膜(mo)(mo)(mo)(mo)(mo)可(ke)(ke)透過PI。例如，認(ren)為(wei)可(ke)(ke)行的(de)(de)(de)(de)細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)是(shi)FITC膜(mo)(mo)(mo)(mo)(mo)聯蛋(dan)(dan)白(bai)(bai)V和(he)PI陰性(xing)(xing)(xing)(xing)(xing); 早(zao)期(qi)(qi)凋(diao)亡(wang)(wang)(wang)(wang)(wang)細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)是(shi)FITC Annexin V陽(yang)(yang)性(xing)(xing)(xing)(xing)(xing)和(he)PI陰性(xing)(xing)(xing)(xing)(xing); 細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)凋(diao)亡(wang)(wang)(wang)(wang)(wang)或已經(jing)(jing)死(si)(si)亡(wang)(wang)(wang)(wang)(wang)的(de)(de)(de)(de)細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)都是(shi)FITC膜(mo)(mo)(mo)(mo)(mo)聯蛋(dan)(dan)白(bai)(bai)V和(he)PI陽(yang)(yang)性(xing)(xing)(xing)(xing)(xing)。該測定(ding)不區分已經(jing)(jing)經(jing)(jing)歷凋(diao)亡(wang)(wang)(wang)(wang)(wang)死(si)(si)亡(wang)(wang)(wang)(wang)(wang)的(de)(de)(de)(de)細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)與由(you)于壞死(si)(si)性(xing)(xing)(xing)(xing)(xing)途徑死(si)(si)亡(wang)(wang)(wang)(wang)(wang)的(de)(de)(de)(de)細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)，因為(wei)在任(ren)一情況下，死(si)(si)細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)都會用FITC Annexin V和(he)PI染(ran)色。然(ran)而，隨著時間的(de)(de)(de)(de)推移，當細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)凋(diao)亡(wang)(wang)(wang)(wang)(wang)被測量時，細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)可(ke)(ke)以經(jing)(jing)常從(cong)FITC膜(mo)(mo)(mo)(mo)(mo)聯蛋(dan)(dan)白(bai)(bai)V和(he)PI陰性(xing)(xing)(xing)(xing)(xing)（可(ke)(ke)行或無可(ke)(ke)測量的(de)(de)(de)(de)凋(diao)亡(wang)(wang)(wang)(wang)(wang)）追溯到(dao)FITC膜(mo)(mo)(mo)(mo)(mo)聯蛋(dan)(dan)白(bai)(bai)V陽(yang)(yang)性(xing)(xing)(xing)(xing)(xing)和(he)PI陰性(xing)(xing)(xing)(xing)(xing)（早(zao)期(qi)(qi)凋(diao)亡(wang)(wang)(wang)(wang)(wang)，膜(mo)(mo)(mo)(mo)(mo)完(wan)整性(xing)(xing)(xing)(xing)(xing)存在），zui后到(dao)FITC膜(mo)(mo)(mo)(mo)(mo)聯蛋(dan)(dan)白(bai)(bai)V和(he)PI陽(yang)(yang)性(xing)(xing)(xing)(xing)(xing)（末期(qi)(qi)凋(diao)亡(wang)(wang)(wang)(wang)(wang)和(he)死(si)(si)亡(wang)(wang)(wang)(wang)(wang)）。細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)通過這三個階(jie)段的(de)(de)(de)(de)運動表明細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)凋(diao)亡(wang)(wang)(wang)(wang)(wang)。相比之下，一個單一的(de)(de)(de)(de)觀察結(jie)果表明，細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)都是(shi)FITC膜(mo)(mo)(mo)(mo)(mo)聯蛋(dan)(dan)白(bai)(bai)V和(he)PI陽(yang)(yang)性(xing)(xing)(xing)(xing)(xing)，本(ben)身就顯示出關于細(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)進行死(si)(si)亡(wang)(wang)(wang)(wang)(wang)的(de)(de)(de)(de)過程的(de)(de)(de)(de)信息(xi)較少。

準備和儲存

在(zai)4°C下(xia)未經稀(xi)釋保存，防止長(chang)時間暴露于光線下(xia)。不要凍結

產品通知

1. 由于應用程序不同，每個研究者應滴定試劑以獲得*結果。
2. 所有血清蛋白的來源均來自美國農業部檢查的位于美國的屠宰場。
3. 注意：疊氮鈉在酸性條件下會產生高毒性的疊氮酸。在棄置之前將水中的疊氮化合物稀釋，以避免在管道中累積潛在的爆炸性沉積物。

DESCRIPTION

Apoptosis is a normal physiologic process which occurs during embryonic development as well as in maintenence of tissue homeostasis. The apoptotic program is characterized by certain morphologic features, including loss of plasma membrane asymmetry and attachment, condensation of the cytoplasm and nucleus, and internucleosomal cleavage of DNA. Loss of plasma membrane is one of the earliest features. In apoptotic cells, the membrane phospholipid phosphatidylserine (PS) is translocated from the inner to the outer leaflet of the plasma membrane, thereby exposing PS to the external cellular environment. Annexin V is a 35-36 kDa Ca2+ dependent phospholipid-binding protein that has a high affinity for PS, and binds to cells with exposed PS. Annexin V may be conjugated to fluorochromes including FITC. This format retains its high affinity for PS and thus serves as a sensitive probe for flow cytometric analysis of cells that are undergoing apoptosis. Since externalization of PS occurs in the earlier stages of apoptosis, FITC Annexin V staining can identify apoptosis at an earlier stage than assays based on nuclear changes such as DNA fragmentation.

FITC Annexin V staining precedes the loss of membrane integrity which accompanies the latest stages of cell death resulting from either apoptotic or necrotic processes. Therefore, staining with FITC Annexin V is typically used in conjunction with a vital dye such as propidium iodide (PI) or 7-Amino-Actinomycin (7-AAD) to allow the investigator to identify early apoptotic cells (PI negative, FITC Annexin V positive). Viable cells with intact membranes exclude PI, wheras the membranes of dead and damaged cells are permeable to PI. For example, cells that are considered viable are FITC Annexin V and PI negative; cells that are in early apoptosis are FITC Annexin V positive and PI negative; and cells that are in late apoptosis or already dead are are both FITC Annexin V and PI positive. This assay does not distinguish between cells that have undergone apoptotic death versus those that have died as a result of a necrotic pathway because in either case, the dead cells will stain with both FITC Annexin V and PI. However, when apoptosis is measured over time, cells can be often tracked from FITC Annexin V and PI negative (viable, or no measurable apoptosis), to FITC Annexin V positive and PI negative (early apoptosis, membrane integrity is present) and finally to FITC Annexin V and PI positive (end stage apoptosis and death). The movement of cells through these three stages suggests apoptosis. In contrast, a single observation indicating that cells are both FITC Annexin V and PI positive, in of itself, reveals less information about the process by which the cells underwent their demise.

FITC膜聯蛋白V細胞凋亡檢測試劑盒（BD Pharmingen）PREPARATION AND STORAGE

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

PRODUCT NOTICES

1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.