Lightning-Link® PE 抗體標定試劑盒

PE / R-Phycoerythrin Conjugation Kit - Lightning-Link®

貨號

包裝

3 x 10 µg
60 µg
3 x 60 µg
600 µg
5 x 600 µg
3 mg

廠牌

產品簡介

Abcam PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® 提供了一種簡便快速的方法，可將 R-藻紅蛋白（R-Phycoerythrin, R-PE，即俗稱的 PE 螢光染料）標定至抗體或其他含有游離胺基的蛋白質、胜肽分子上，標定產物可直接應用於流式細胞分析 (flow cytometry)、ELISA、免疫組織化學染色 (IHC) 等實驗。套組採用 Lightning-Link® 直接標定技術，僅需將修飾劑 (Modifier) 加入待標定抗體，再以此溶液回溶凍乾的 PE 試劑小瓶，即可啟動標定反應；動手操作時間約 30 秒，3.5 小時左右即可完成標定，全程無需任何純化步驟，抗體回收率可達 100%。

主要特色

操作簡便快速，套組採用 Lightning-Link® 直接標定技術，僅需將添加有 Modifier 的待標定抗體與凍乾的 PE 試劑混合，即可透過離胺酸 (lysine) 等游離胺基進行共價標定，動手操作時間僅約 30 秒，整體反應時間在 3.5 小時左右（標定反應約 3 小時＋中止反應 30 分鐘）。
全程無需純化步驟，抗體回收率可達 100%，標定完成後即可直接應用於 Flow cytometry、ELISA、IHC 等實驗，無需額外處理。

Flow Cytometry. Robinson, Andrew P., et al used PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918) as part of characterizing oligodendroglial populations. They used the kit to conjugate PE / R-Phycoerythrin to Mouse monoclonal anti-GALC antibody for use in flow cytometry. SJL/J mice were immunized with PLP139-151 and scored daily for clinical disease. A cohort of SJL/J mice was sacrificed, and spinal cords were analyzed by flow cytometry (n=5). (A) Cells were distinguished from debris by forward and side scatter then singlet cells were gated. Live cells were gated by dead cell exclusion, and CNS resident cells were identified as CD45- or CD45low. (B) Oligodendroglial cells were defined by double positive staining : A2B5+PDGFRalpha+ early OPCs, A2B5+NG2+ intermediate OPCs, NG2+O4+ late OPCs, O4+MOG+ pre-myelinating oligodendrocytes, and GALC+MOG+ mature oligodendrocytes. — **Flow Cytometry.** Robinson, Andrew P., et al used PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918) as part of characterizing oligodendroglial populations. They used the kit to conjugate PE / R-Phycoerythrin to Mouse monoclonal anti-GALC antibody for use in flow cytometry. SJL/J mice were immunized with PLP_139-151 and scored daily for clinical disease. A cohort of SJL/J mice was sacrificed, and spinal cords were analyzed by flow cytometry (n=5). (A) Cells were distinguished from debris by forward and side scatter then singlet cells were gated. Live cells were gated by dead cell exclusion, and CNS resident cells were identified as CD45^- or CD45^low. (B) Oligodendroglial cells were defined by double positive staining : A2B5+PDGFRalpha⁺ early OPCs, A2B5+NG2+ intermediate OPCs, NG2+O4+ late OPCs, O4+MOG+ pre-myelinating oligodendrocytes, and GALC+MOG+ mature oligodendrocytes. *Image from Robinson, Andrew P., et al., PloS one, 9(9):e107649. doi: 10.1371/journal.pone.0107649. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/*

Flow Cytometry. Flow cytometry histogram showing integrin beta-3 positive population of platelets from wildtype and knockout mice. Integrin beta-3 antibody was conjugated using EasyLink R-Phycoerythrin conjugation kit (ab102918). Flow cytometry was performed using platelets from wild type and integrin beta-3 knockout mice. Mice that expressed beta-3 (shown in red) had a clear shift in FL-2 fluorescence over beta-3 knockout mice (shown in black). — **Flow Cytometry.** Flow cytometry histogram showing integrin beta-3 positive population of platelets from wildtype and knockout mice. Integrin beta-3 antibody was conjugated using EasyLink R-Phycoerythrin conjugation kit (ab102918). Flow cytometry was performed using platelets from wild type and integrin beta-3 knockout mice. Mice that expressed beta-3 (shown in red) had a clear shift in FL-2 fluorescence over beta-3 knockout mice (shown in black). *This image is courtesy of an anonymous Abreview.*

Microsphere immunoassay based on Luminex. Charlermroj R et al. used ab102918 as part of an experiment studying four different plant pathogens : a fruit blotch bacterium Acidovorax avenae subsp. citrulli (Aac), chilli vein-banding mottle virus (CVbMV, potyvirus), watermelon silver mottle virus (WSMoV, tospovirus serogroup IV) and melon yellow spot virus (MYSV, tospovirus). (A-G) X-axis is antibody-coated microsphere and y-axis is median fluorescent intensity (MFI) from each RPE-labeled antibody. (H) Summary of selected antibody pair sets for the detection of the four plant pathogens. — **Microsphere immunoassay based on Luminex.** Charlermroj R et al. used ab102918 as part of an experiment studying four different plant pathogens : a fruit blotch bacterium Acidovorax avenae subsp. citrulli (Aac), chilli vein-banding mottle virus (CVbMV, potyvirus), watermelon silver mottle virus (WSMoV, tospovirus serogroup IV) and melon yellow spot virus (MYSV, tospovirus). (A-G) X-axis is antibody-coated microsphere and y-axis is median fluorescent intensity (MFI) from each RPE-labeled antibody. (H) Summary of selected antibody pair sets for the detection of the four plant pathogens. *Image from Charlermroj R et al., PloS One, 8(4):e62344. Fig 2.; doi: 10.1371/journal.pone.0062344. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/*

實驗注意事項

樣本緩衝液 pH 值須介於 6.5–8.5，相容成分包括：Tris（濃度應 ≤50 mM / 0.6%）、BSA（≤0.1%）、甘油 (glycerol，≤50%)、疊氮化鈉 (Sodium Azide，≤0.1%)、磷酸鉀、氯化鈉、蔗糖、檸檬酸鈉、海藻糖等。緩衝液不可含有 Thimerosal (Thiomersal)、Proclin、Glycine、Arginine、Glutathione 等成分，以免干擾標定反應。更多樣本緩衝液細部條件說明，可參照下方表格或產品說明書。
僅適用於已純化的抗體樣本；若樣本中含有其他蛋白質、胜肽或游離胺基分子，可能會競爭消耗標定試劑，影響標定效率。抗體樣本理想濃度為 1 mg/mL，若濃度不足（< 0.5 mg/ml）或過高（> 1 mg/ml），建議先進行濃縮或稀釋處理。
標定完成的產物於 4°C 保存可穩定達 18 個月；若需長期保存，建議加入 50% 甘油作為冷凍保護劑，並存放於 -20°C。實際最適合的保存條件仍視個別抗體的穩定性而定，建議透過實驗確認。

Compatible buffer constituents
50mM / 0.6% Tris¹	0.1% BSA	50% glycerol
0.1% sodium azide	PBS	Potassium phosphate
Sodium chloride	HEPES	Sucrose
Sodium citrate	EDTA	Trehalose

¹ Tris buffered saline is almost always ≤ 50 mM / 0.6%

Incompatible buffer constituents
Thiomerosal	Proclin	Glycine
Arginine	Glutathione	DTT

套組內容物

Item	3 x 10 µg	60 µg	3 x 60 µg	600 µg	5 x 600 µg	3 mg	Storage temperature
PE / R- Phycoerythrin mix	3 vials	1 vial	3 vials	1 vial	5 vials	1 vial	-20°C
Modifier reagent	1 vial	1 vial	1 vial	1 vial	1 vial	1 vial	-20°C
Quencher reagent	1 vial	1 vial	1 vial	1 vial	1 vial	1 vial	-20°C

產品說明書

ab102918 PE / R-Phycoerythrin Conjugation Kit - Lightning-Link®