新品｜细胞内流式细胞术 Intracellular flow cytometry
新品介紹 — 细胞内流式细胞术
Intracellular Flow Cytometry
為我們提供了一種強大的工具來對細胞內的目標蛋白進行染色，例如：磷酸化訊號轉導(phosphorylated signal transducers)、轉錄因子(transcription factors)和細胞因子(cytokines)。通過固定和透化的步驟，抗體能夠穿透細胞膜和核膜對細胞內的目標蛋白染色。在這方面，Proteintech 提供一系列使用於單細胞分析的細胞內流式細胞抗體，除此之外，在固定劑和透化劑的部分也經過了優化，以讓您的實驗獲得可重複的結果。
关於细胞内的目标在这裡可分為两个部分：细胞质(颁测迟辞辫濒补蝉尘颈肠)以及细胞核(狈耻肠濒别补谤)。
细胞质目标检测(Cytoplasmic Targets Detection)
细胞内流式细胞术是一種強大的技術，可以同時檢測單個細胞內的多項目標。细胞内流式细胞术也是分析免疫或發炎反應過程中 T 細胞、B 細胞、NK 細胞分化、活化和可塑性的有效工具。
细胞质目标中较常使用到的是细胞因子(颁测迟辞办颈苍别) 的探測，即免疫和發炎反應相關的分泌因子。在細胞內流式細胞技術中，透過蛋白質轉運抑製劑(protein transport inhibitors) 的使用，例如： Brefeldin A 或monensin，可將細胞因子捕獲在分泌細胞內，以實現在異質群體(heterogenous population)中精準定位特定細胞因子的來源。除了細胞因子，其他細胞質蛋白也可以通過此技術檢測到。在這方面，Proteintech 提供各種螢光染料標記的抗體，涵蓋了大量的目標和不同顏色，以及相關試劑，以供您做選擇。
除了細胞因子，其他細胞質蛋白也可以通過细胞内流式细胞术檢測到。Proteintech 提供各種螢光染料標記的抗體，涵蓋大量目標和不同顏色，以及用於細胞質目標細胞內流動分析的相關試劑。
以下是实验结果展示：
▲Human PBMCs are treated by LPS and protein transporter Brefeldin A. 1*10^6 stimulated human PBMCs were surface stained with (, Clone: UCHM-1), fixed by 4% PFA and then permeabilized by (). Cells were then intracellularly stained with 1ul (, Clone: 4G3E6), or .
▲Human PBMCs are stimulated by PHA for 3 days. Cells were intracellularly stained with 5ul (red), or 0.2ug (blue). Cells were fixed by 4% PFA and permeabilized with (). Lymphocytes are gated.
細胞質目標(Cytoplasmic Targets)相關抗體
| Cat No. | Product Name | Clone No. | Host | Reactivity |
| CoraLite®488-conjugated GAPDH Monoclonal antibody | 1E6D9 | Mouse | Human, Mouse, Rat, Zebrafish, Yeast, Plant | |
| CoraLite®488-conjugated BAX Monoclonal antibody | 4G5E8 | Mouse | Human, Mouse, Rat | |
| CoraLite®488-conjugated Bcl-XL Monoclonal antibody | 4C12A6 | Mouse | Human, Dog | |
| CoraLite®488-conjugated IGF2R-Specific Polyclonal antibody | Rabbit | Human | ||
| CoraLite®488-conjugated Osteopontin Polyclonal antibody | Rabbit | Human, Rat, Mouse | ||
| CoraLite®594-conjugated Galectin-3 Monoclonal antibody | 1C1B2 | Mouse | Human, Mouse, Rat | |
| CoraLite®594-conjugated SLP76 Monoclonal antibody | 4H10B1 | Mouse | Human, Dog | |
| CoraLite®594-conjugated VEGFA Polyclonal antibody | Rabbit | Human | ||
| CoraLite®594-conjugated 14-3-3 Sigma Monoclonal antibody | 3F10D9 | Mouse | Human | |
| CoraLite®594-conjugated Albumin Monoclonal antibody | 4A1C11 | Mouse | Human, Rat, Pig | |
| CoraLite®647-conjugated BMP-7 Monoclonal antibody | 3E9G7 | Mouse | Human | |
| CoraLite®647-conjugated IL-6 Monoclonal antibody | 4G3E6 | Mouse | Human | |
| CoraLite®647-conjugated CPT1A Monoclonal antibody | 5D8G9 | Mouse | Human, Mouse, Rat | |
| CoraLite®647-conjugated PDI Polyclonal antibody | Rabbit | Human, Mouse, Rat | ||
| CoraLite®647-conjugated TGF beta 1 Monoclonal antibody | 1E3E9 | Mouse | Human | |
| APC Anti-Human CD68 (Y1/82A) | Y1/82A | Mouse | Human | |
| APC Anti-Mouse CD107b (ABL-93) | ABL-93 | Rat | Mouse | |
| FITC-conjugated ATF4 Polyclonal antibody | Rabbit | Human, Mouse | ||
| FITC Anti-Mouse CD107a (1D4B) | 1D4B | Rat | Mouse | |
| PE Anti-Mouse IFN gamma (XMG1.2) | XMG1.2 | Rat | Mouse | |
| PE Anti-Mouse IL-12/IL-23 p40 (C17.8) | C17.8 | Rat | Mouse |
细胞核目标检测(Nuclear Targets Detection)
實驗步驟與细胞质目标检测類似，轉錄因子和其他核蛋白的染色需要在表面標記(Surface marker)染色後進行細胞固定和透化。而根據轉錄因子的性質以及在細胞核、DNA 或其他蛋白質上的位置，可能會需要不同的固定和透化步驟。
细胞的固定和透化步骤可能会影响到细胞表面标记的染色，因此对於缓衝液的选择便顯得尤為重要。為了讓大家可以輕鬆的操作轉錄因子檢測，在這裡Proteintech 建議使用，这个是專門為使用流式細胞儀分析核目標時，所能達成的最佳分辨率和低背景而設計的。通過试剂盒中的固定和透化試劑，抗體能夠穿透細胞和核膜，對細胞內的目標蛋白進行染色。結合Proteintech精選的细胞内流式细胞术抗體，該试剂盒將會為您帶來更好的細胞內流式細胞檢測結果！
以下是实验结果展示：
▲1X10^6 mouse splenocytes were surface stained with (, Clone: GK1.5) and then fixed by (). Cells were then stained with 5 ul (, Clone: 3G3), or .
細胞核目標(Nuclear Targets)相關抗體
| Cat No. | Product Name | Clone No. | Host | Reactivity |
| CoraLite®488-conjugated TDP-43 (C-terminal) Polyclonal antibody | Rabbit | Human, Mouse, Rat | ||
| CoraLite®488-conjugated c-MYC Monoclonal antibody | 3D9C12 | Mouse | Human, Mouse, Rat | |
| CoraLite®488-conjugated HDAC2 Polyclonal antibody | Rabbit | Human, Mouse, Rat | ||
| CoraLite®488-conjugated NeuN Monoclonal antibody | 3A4C1 | Mouse | Human, Mouse, Rat | |
| CoraLite®594-conjugated SOX2 Monoclonal antibody | 4B9B10 | Mouse | Human, Pig, Mouse | |
| CoraLite®594-conjugated PAX8 Polyclonal antibody | Rabbit | Human, Mouse, Rat | ||
| CoraLite®594-conjugated CCAR2 Monoclonal antibody | 2D10F8 | Mouse | Human, Pig | |
| CoraLite®594-conjugated NRF2, NFE2L2 Polyclonal antibody | Rabbit | Human, Mouse, Rat | ||
| CoraLite®647-conjugated EED Monoclonal antibody | 1D11D4 | Mouse | Human, Mouse | |
| CoraLite®647-conjugated RBPMS Polyclonal antibody | Rabbit | Human, Mouse, Rat | ||
| CoraLite®647-conjugated Lamin B1 Monoclonal antibody | 3C10G12 | Mouse | Human, Mouse, Rat | |
| APC Anti-Mouse Foxp3 (3G3) | 3G3 | Mouse | Mouse | |
| PE Anti-Mouse Foxp3 (3G3) | 3G3 | Mouse | Mouse |
其他相关试剂产物
(Cat No. )
Foxp3 / Transcription Factor Staining Buffer Kit中包含專門配製的緩衝液和試劑，讓您通過流式細胞術分析核抗原時，能夠獲得最佳分辨率和低背景。下表為试剂盒中所附的試劑，可在檢測Foxp3 和 ROR gamma等核抗原的染色實驗中使用。
| Components | Cat No. | Size |
| Foxp3 / Transcription Factor Fix/Perm Concentrate (4X) | 50 mL | |
| Foxp3 / Transcription Factor Fix/Perm Diluent (1X) | 150 mL | |
| Flow Cytometry Perm Buffer (10X) | 150 mL |
(Cat No. )
Flow Staining Buffer 是一種緩衝的生理食鹽水溶液，其中含有小牛血清作為蛋白質來源，以最大程度地減少抗體和疊氮化鈉 (0.09%) 的非特異性結合作為防腐劑。
此缓衝液适用於稀释抗体、细胞悬浮液，以及作為细胞表面染色和流式细胞分析过程的洗滌緩衝液(wash buffer)。
(Cat No. )
RBC Lysis Buffer (10X) 是一種濃縮的氯化銨 (ammonium chloride) 裂解試劑。 可用於全血样品的预处理，用來溶解單细胞悬浮液中的紅血球，對白血球的影響最小。另外，RBC Lysis Buffer (10X) 中不含固定剂，因此细胞在使用过后仍能存活。
Isotype control
實驗當中擁有一個Control 組別是一件非常重要的事情，能夠幫助判讀實驗的誤差，以及判斷樣品和試劑是否有問題。因此在這裡我們也提供了適用於Flow cytometry的相關Isotype control。
| Cat No. | Ig Class/Subclass | Species | Isotype Control |
| IgG | Rabbit | ||
| IgG1 | Mouse | ||
| IgG2a | Mouse | ||
| IgG2b | Mouse | ||
| IgG1 | Rat | ||
| IgG2a | Rat | ||
| IgG2b | Rat |
其他重要资讯
1. 緩衝液及溶液的製備
2. 實驗步驟
