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.
Components
Part ? No.
Description
Size
51-65874X
FITC ? Annexin V
0.5 ? mL (1 ea)
51-66211E
Propidium ? Iodide Staining Solution
2 ? mL (1 ea)
51-66121E
10X ? Annexin V Binding Buffer
50 ? mL (1 ea)
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.
4.Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
]]>BD 555899 Lysing Buffer 紅細(xì)胞裂解液
產(chǎn)品品牌:BD
產(chǎn)品產(chǎn)地:美國(guó)
產(chǎn)品規(guī)格:100ml
用途范圍:流式細(xì)胞術(shù)(常規(guī)檢測(cè))
BD Pharm Lyse
lysing solution is a buffered, concentrated (10X) ammonium chloride-based lysing reagent. When diluted to a 1X concentration and used as recommended, BD Pharm Lyse lyses red blood cells following monoclonal antibody staining. The lysing solution results in good light scatter separation of lymphocytes and red blood cell debris when analyzed by flow cytometry.??BD Pharm Lyse does not contain a fixative agent, so leukocytes remain viable after red blood cell lysis.
BD Pharm Lyse 裂解液是一種緩沖、濃縮 (10X) 氯化銨基裂解試劑。 當(dāng)稀釋至 1X 濃度并按照推薦使用時(shí)，BD Pharm Lyse 會(huì)在單克隆抗體染色后裂解紅細(xì)胞。 當(dāng)通過(guò)流式細(xì)胞儀分析時(shí)，裂解溶液導(dǎo)致淋巴細(xì)胞和紅細(xì)胞碎片的良好光散射分離。 BD Pharm Lyse 不含固定劑，因此白細(xì)胞在紅細(xì)胞裂解后仍可存活。
品牌：BD Pharm Lyse
監(jiān)管狀態(tài)：RUO
RRID：AB_2869057
應(yīng)用：流式細(xì)胞儀（常規(guī)測(cè)試）
1X diluted solutions may be stored at 4° C for up to 30 days.
Preparation of 1X lysing solution:??Dilute the 10X concentrate 1:10 with distilled water. The pH of the 1X solution should fall within the range of pH 7.1-7.4.??Adjust the pH if necessary. Warm the 1X solution to room temperature prior to use. 100 ml of 10X concentrate will yield a quantity of 1X solution that is sufficient to lyse 500 samples.
Lysing procedure:
Note: The following procedure is only applied to human whole blood red blood cell lysis. Since applications vary, for other samples such as mouse spleen red blood cell lysis (lysing incubation time up to 3 minutes at 37°C), bone marrow red blood cell lysis, each investigator should optimize the condition to obtain optimal results.
1. Add 2.0 ml??of 1X lysing solution to each tube containing up to 200 μl of a whole blood plus monoclonal antibody mixture.
2. Gently vortex each tube immediately after adding the lysing solution.
3. Incubate at room temperature, protected from light, for 15 minutes.
4. Centrifuge 200 X g for 5 minutes.
5. Carefully aspirate supernatent, without disturbing pellet.
6. Add 2.0 ml 1X PBS containing 1% heat-inactivated fetal bovine serum and 0.1% sodium azide (PBS-FBS).
7. Centrifuge at 200 X g for 5 minutes.
8. Carefully aspirate supernatent, without disturbing pellet.
9. Resuspend pellet in 0.5 ml PBS-FBS or a fixative such as 2% formaldehyde for flow cytometric analysis.
Troubleshooting:?Incomplete lysis may occur for several reasons: (1) The age of the specimen may affect red cell lysis, (2) The 1X lysing solution may not have been warmed to room temperature, or (3) After adding the lysing solution, the samples may not have been vortexed sufficiently. If incomplete lysis occurs (recognized by the presence of an excessive amount of visible red blood cells in the final, washed cell suspension), repeat lysing procedure. However, lysing more than two times is not recommended.
產(chǎn)品簡(jiǎn)介:
BD Pharm Lyse裂解液是一種緩沖的,濃縮的(10倍)lv hua 銨基裂解試劑。當(dāng)稀釋到1X濃度并按推薦使用時(shí),BD Pharm Lyse在單克隆抗體染色后溶解紅細(xì)胞。當(dāng)用流式細(xì)胞術(shù)分析時(shí),溶解液能很好的光散射分離淋巴細(xì)胞和紅細(xì)胞碎片。BD Pharm Lyse不含固定劑,所以白細(xì)胞在紅細(xì)胞溶解后仍可存活。
產(chǎn)品應(yīng)用:
流式細(xì)胞術(shù)(常規(guī)檢測(cè))
運(yùn)輸與儲(chǔ)存:
保存在4°C未稀釋。
1X稀釋的溶液可在4°C下保存30天。
推薦的實(shí)驗(yàn)流程:
1X溶出液的制備:將10X濃縮液以1:10的比例用蒸餾水稀釋。1X溶液的pH值應(yīng)在pH 7.1-7.4范圍內(nèi)。必要時(shí)調(diào)整pH值。使用前將1X溶液加熱至室溫。100ml的10X濃縮液將產(chǎn)生足夠溶解500個(gè)樣品的1X溶液。
溶解過(guò)程:
注:以下步驟僅適用于人全血紅細(xì)胞裂解。由于應(yīng)用的不同,對(duì)于其他樣品,如小鼠pi zang紅細(xì)胞裂解(在37℃下裂解培養(yǎng)時(shí)間可達(dá)3分鐘),gu sui紅細(xì)胞裂解,每個(gè)研究人員應(yīng)優(yōu)化條件,以獲得 結(jié)果。
3.室溫避光孵育15分鐘。
產(chǎn)品訂購(gòu)信息:
貨號(hào)? ? ? ? ? ? ? ? 名稱? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?規(guī)格
555899? ? Lysing Buffer 10X Concentrate? ? ? 100mL
BD 349202 FACS Lysing Solution
BD FACS lysing solution is intended for lysing red blood cells following direct immunofluorescence staining of human peripheral blood cells with monoclonal antibodies prior to flow cytometric analysis.
BD FACS lysing solution is appropriate for use with reagents such as BD Tritest or BD Simultest reagents and a suitably equipped flow cytometer. It may be used in both lyse/wash and lyse/no-wash procedures.
BD FACS 裂解液用于在流式細(xì)胞術(shù)分析之前用單克隆抗體對(duì)人外周血細(xì)胞進(jìn)行直接免疫熒光染色后裂解紅細(xì)胞。
BD FACS 裂解液適用于試劑，例如 BD Tritest 或 BD Simultest 試劑和適當(dāng)配備的流式細(xì)胞儀。 它可用于裂解/洗滌和裂解/免洗滌程序。
反應(yīng)性：人類
存儲(chǔ)緩沖區(qū)：含有 <15% 甲醛和 <50% 二甘醇的緩沖溶液。
應(yīng)用：流式細(xì)胞儀
BD FACS lysing solution is intended for lysing red blood cells following direct immunofluorescence staining of human peripheral blood cells with monoclonal antibodies prior to flow cytometric analysis.
BD FACS lysing solution is appropriate for use with reagents such as BD Tritest or BD Simultest reagents ａnd a suitably equipped flow cytometer. It may be used in both lyse/wash ａnd lyse/no-wash procedures
FACS溶血素（BDIS Catalog No.349202）用于PBMCs，培養(yǎng)細(xì)胞與全血制備，主要有3個(gè)作用：
(1) 全血制備時(shí)用于溶解紅細(xì)胞
(2) 固定外表位
(3) 輔助通透劑
???? FACS溶血素使用前用無(wú)離子水1：10稀釋，勿用PBS或其它緩沖液。而且FACS溶血素必須與FACS通透液一起使用，即使是培養(yǎng)細(xì)胞的檢測(cè)。