新品介紹 — 細胞內流式細胞術
Intracellular Flow Cytometry
細胞內流式細胞術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。
關於細胞內的目標在這裡可分為兩個部分：細胞質(Cytoplasmic)以及細胞核(Nuclear)。
細胞質目標檢測(Cytoplasmic Targets Detection)
細胞內流式細胞術是一種強大的技術，可以同時檢測單個細胞內的多項目標。細胞內流式細胞術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。
細胞質目標中較常使用到的是細胞因子(Cytokine) 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細胞內流式細胞術檢測到。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。
以下是實驗結果展示：
▲Human PBMCs are treated by LPS and protein transporter Brefeldin A. 1*10^6 stimulated human PBMCs were surface stained with PE Anti-human CD14 (PE-65056, Clone: UCHM-1), fixed by 4% PFA and then permeabilized by 1X Flow Cytometry permeabilization buffer (PF00011-C). Cells were then intracellularly stained with 1ul CoraLite®647-conjugated Anti-human IL6 (CL647-69001, Clone: 4G3E6), or CoraLite®647-conjugated mouse IgG1 isotype control.
▲Human PBMCs are stimulated by PHA for 3 days. Cells were intracellularly stained with 5ul CoraLite488 Anti-human CD107a (H4A3) (red), or 0.2ug CoraLite488 mouse IgG1 isotype control (blue). Cells were fixed by 4% PFA and permeabilized with 1X intracellular flow cytometry perm buffer (PF00011-C). Lymphocytes are gated.
細胞質目標(Cytoplasmic Targets)相關抗體
| Cat No. | Product Name | Clone No. | Host | Reactivity |
| CL488-60004 | CoraLite®488-conjugated GAPDH Monoclonal antibody | 1E6D9 | Mouse | Human, Mouse, Rat, Zebrafish, Yeast, Plant |
| CL488-60267 | CoraLite®488-conjugated BAX Monoclonal antibody | 4G5E8 | Mouse | Human, Mouse, Rat |
| CL488-66020 | CoraLite®488-conjugated Bcl-XL Monoclonal antibody | 4C12A6 | Mouse | Human, Dog |
| CL488-20253 | CoraLite®488-conjugated IGF2R-Specific Polyclonal antibody | Rabbit | Human | |
| CL488-22952 | CoraLite®488-conjugated Osteopontin Polyclonal antibody | Rabbit | Human, Rat, Mouse | |
| CL594-60207 | CoraLite®594-conjugated Galectin-3 Monoclonal antibody | 1C1B2 | Mouse | Human, Mouse, Rat |
| CL594-66465 | CoraLite®594-conjugated SLP76 Monoclonal antibody | 4H10B1 | Mouse | Human, Dog |
| CL594-19003 | CoraLite®594-conjugated VEGFA Polyclonal antibody | Rabbit | Human | |
| CL594-66251 | CoraLite®594-conjugated 14-3-3 Sigma Monoclonal antibody | 3F10D9 | Mouse | Human |
| CL594-66051 | CoraLite®594-conjugated Albumin Monoclonal antibody | 4A1C11 | Mouse | Human, Rat, Pig |
| CL647-69011 | CoraLite®647-conjugated BMP-7 Monoclonal antibody | 3E9G7 | Mouse | Human |
| CL647-69001 | CoraLite®647-conjugated IL-6 Monoclonal antibody | 4G3E6 | Mouse | Human |
| CL647-66039 | CoraLite®647-conjugated CPT1A Monoclonal antibody | 5D8G9 | Mouse | Human, Mouse, Rat |
| CL647-11245 | CoraLite®647-conjugated PDI Polyclonal antibody | Rabbit | Human, Mouse, Rat | |
| CL647-69012 | CoraLite®647-conjugated TGF beta 1 Monoclonal antibody | 1E3E9 | Mouse | Human |
| APC-65187 | APC Anti-Human CD68 (Y1/82A) | Y1/82A | Mouse | Human |
| APC-65052 | APC Anti-Mouse CD107b (ABL-93) | ABL-93 | Rat | Mouse |
| FITC-10835 | FITC-conjugated ATF4 Polyclonal antibody | Rabbit | Human, Mouse | |
| FITC-65050 | FITC Anti-Mouse CD107a (1D4B) | 1D4B | Rat | Mouse |
| PE-65153 | PE Anti-Mouse IFN gamma (XMG1.2) | XMG1.2 | Rat | Mouse |
| PE-65097 | PE Anti-Mouse IL-12/IL-23 p40 (C17.8) | C17.8 | Rat | Mouse |
細胞核目標檢測(Nuclear Targets Detection)
實驗步驟與細胞質目標檢測類似，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。
細胞的固定和透化步驟可能會影響到細胞表面標記的染色，因此對於緩衝液的選擇便顯得尤為重要。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 Foxp3 / Transcription Factor Staining Buffer Kit (Catalog #PF00011)，這個試劑盒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。通過試劑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精選的細胞內流式細胞術抗體，該試劑盒將會為您帶來更好的細胞內流式細胞檢測結果！
以下是實驗結果展示：
▲1X10^6 mouse splenocytes were surface stained with PE Anti-Mouse CD4 (GK1.5) (PE-65104, Clone: GK1.5) and then fixed by Transcription Factor fixation buffer and permeabilization buffer (PF00011-C). Cells were then stained with 5 ul CoraLite®647-conjugated Anti-Mouse Foxp3 (CL647-65089, Clone: 3G3), or CoraLite®647-conjugated mouse IgG1 isotype control.
細胞核目標(Nuclear Targets)相關抗體
| Cat No. | Product Name | Clone No. | Host | Reactivity |
| CL488-12892 | CoraLite®488-conjugated TDP-43 (C-terminal) Polyclonal antibody | Rabbit | Human, Mouse, Rat | |
| CL488-67447 | CoraLite®488-conjugated c-MYC Monoclonal antibody | 3D9C12 | Mouse | Human, Mouse, Rat |
| CL488-12922 | CoraLite®488-conjugated HDAC2 Polyclonal antibody | Rabbit | Human, Mouse, Rat | |
| CL488-66836 | CoraLite®488-conjugated NeuN Monoclonal antibody | 3A4C1 | Mouse | Human, Mouse, Rat |
| CL594-66411 | CoraLite®594-conjugated SOX2 Monoclonal antibody | 4B9B10 | Mouse | Human, Pig, Mouse |
| CL594-10336 | CoraLite®594-conjugated PAX8 Polyclonal antibody | Rabbit | Human, Mouse, Rat | |
| CL594-66497 | CoraLite®594-conjugated CCAR2 Monoclonal antibody | 2D10F8 | Mouse | Human, Pig |
| CL594-16396 | CoraLite®594-conjugated NRF2, NFE2L2 Polyclonal antibody | Rabbit | Human, Mouse, Rat | |
| CL647-67756 | CoraLite®647-conjugated EED Monoclonal antibody | 1D11D4 | Mouse | Human, Mouse |
| CL647-15187 | CoraLite®647-conjugated RBPMS Polyclonal antibody | Rabbit | Human, Mouse, Rat | |
| CL647-66095 | CoraLite®647-conjugated Lamin B1 Monoclonal antibody | 3C10G12 | Mouse | Human, Mouse, Rat |
| APC-65089 | APC Anti-Mouse Foxp3 (3G3) | 3G3 | Mouse | Mouse |
| PE-65089 | PE Anti-Mouse Foxp3 (3G3) | 3G3 | Mouse | Mouse |
其他相關試劑產品
Foxp3 / Transcription Factor Staining Buffer Kit (Cat No. PF00011)
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。下表為試劑盒中所附的試劑，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) | PF00011-A | 50 mL |
| Foxp3 / Transcription Factor Fix/Perm Diluent (1X) | PF00011-B | 150 mL |
| Flow Cytometry Perm Buffer (10X) | PF00011-C | 150 mL |
Flow Cytometry Staining Buffer (1X) (Cat No. PF00012)
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。
此緩衝液適用於稀釋抗體、細胞懸浮液，以及作為細胞表面染色和流式細胞分析過程的洗滌緩衝液(wash buffer)。
RBC Lysis Buffer (10X) (Cat No. PF00013)
RBC Lysis Buffer (10X) 是一種濃縮的氯化銨 (ammonium chloride) 裂解試劑。 可用於全血樣品的預處理，用來溶解單細胞懸浮液中的紅血球，對白血球的影響最小。另外，RBC Lysis Buffer (10X) 中不含固定劑，因此細胞在使用過後仍能存活。
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 |
| 30000-0-AP | IgG | Rabbit | IgG control Polyclonal antibody |
| 66360-1-Ig | IgG1 | Mouse | Mouse IgG1 isotype control Monoclonal antibody |
| 66360-2-Ig | IgG2a | Mouse | Mouse IgG2a isotype control Monoclonal antibody |
| 66360-3-Ig | IgG2b | Mouse | Mouse IgG2b isotype control Monoclonal antibody |
| CL647-65212 | IgG1 | Rat | CoraLite®647 Rat IgG1 Isotype Control (HRPN) |
| CL647-65209 | IgG2a | Rat | CoraLite®647 Rat IgG2a Isotype Control (2A3) |
| CL647-65211 | IgG2b | Rat | CoraLite®647 Rat IgG2b Isotype Control (LTF-2) |
其他重要資訊
1. 緩衝液及溶液的製備Buffer and Solution Preperation
2. 實驗步驟Experimental Procedure
