Lightning-Link® APC 抗體標定試劑盒

APC Conjugation Kit - Lightning-Link®

貨號

ab201807

包裝

3 x 10 μg
100 μg
3 x 100 μg
1 mg

廠牌

Abcam

產品簡介

Abcam APC Conjugation Kit - Lightning-Link® 提供了一種簡便快速的方法，可將 APC 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 Western Blot、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 APC 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%。

Abcam Lightning-Link® Conjugation Kit

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 APC 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 Western Blot、Flow cytometry、ELISA、IHC 等實驗，無需額外處理。
Fluorescence Microscopy. Haque, Nasreen S., Akaash Tuteja, and Niloufar Haque used APC Conjugation Kit - Lightning-Link® (ab201807) as part of examining as part of examining embryoid body formation. They used the kit to conjugate APC to anti-CCR8 antibody for use in immunofluorescence. Anti CCL1 inhibits the expression of CCR8 and FoxP3 in mouse mesenchymal stem cells (mMSCs). Untreated (5a,-c) and anti-CCL1 treated (5d,-,f) cells were subjected to immunoflourescence with antibodies against FoxP3 (green;5b and e) or CCR8 (red;5c and f).
Fluorescence Microscopy. Haque, Nasreen S., Akaash Tuteja, and Niloufar Haque used APC Conjugation Kit - Lightning-Link® (ab201807) as part of examining as part of examining embryoid body formation. They used the kit to conjugate APC to anti-CCR8 antibody for use in immunofluorescence. Anti CCL1 inhibits the expression of CCR8 and FoxP3 in mouse mesenchymal stem cells (mMSCs). Untreated (5a,-c) and anti-CCL1 treated (5d,-,f) cells were subjected to immunoflourescence with antibodies against FoxP3 (green;5b and e) or CCR8 (red;5c and f). Image from Haque, Nasreen S., Akaash Tuteja, and Niloufar Haque., PloS one 14.7 (2019): e0218944. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Fluorescence Microscopy. Kim, Nayoung, et al used APC Conjugation Kit - Lightning-Link® (ab201807) as part of examining apoptosis in HIV-1-infected CD4+ primary T cells. They used the kit to conjugate APC to anti-pFOXO3a antibody for use in confocal microscopy. Jurkat T cells expressing tTA alone, TatSF2+tTA, or TatSF2G48-R57A +tTA were stained with antibodies against PPP2R1B (first and forth columns of panels, green), pFOXO3a (second column, red), and FOXO3a (forth column, red).
Fluorescence Microscopy. Kim, Nayoung, et al used APC Conjugation Kit - Lightning-Link® (ab201807) as part of examining apoptosis in HIV-1-infected CD4+ primary T cells. They used the kit to conjugate APC to anti-pFOXO3a antibody for use in confocal microscopy. Jurkat T cells expressing tTA alone, TatSF2+tTA, or TatSF2G48-R57A +tTA were stained with antibodies against PPP2R1B (first and forth columns of panels, green), pFOXO3a (second column, red), and FOXO3a (forth column, red). Image from Kim, Nayoung, et al., PLoS Pathog., 6(9): e1001103. doi: 10.1371/journal.ppat.1001103. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Flow Cytometry. Robinson, Andrew P., et al used APC Conjugation Kit - Lightning-Link® (ab201807) as part of characterizing oligodendroglial populations. They used the kit to conjugate APC to Mouse anti-O4 antibody, clone 81, 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+PDGFRα+ 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 APC Conjugation Kit - Lightning-Link® (ab201807) as part of characterizing oligodendroglial populations. They used the kit to conjugate APC to Mouse anti-O4 antibody, clone 81, 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+PDGFRα+ 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/

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% Tris10.1% BSA250% glycerol
0.1% sodium azidePBSPotassium phosphate
Sodium chlorideHEPESSucrose
Sodium citrateEDTATrehalose
1 Tris buffered saline is almost always ≤ 50 mM / 0.6%
2 BSA can also interfere with the use of the conjugated antibody in tissue staining.
Incompatible buffer constituents
ThiomerosalProclinGlycine
ArginineGlutathioneDTT

套組內容物

Item3 x 10 µg100 µg3 x 100 µg1 mgStorage temperature
APC mix3 vials1 vial3 vials1 vial-20°C
Modifier reagent1 vial1 vial1 vial1 vial-20°C
Quencher reagent1 vial1 vial1 vial1 vial-20°C

產品說明書

ab201807 APC Conjugation Kit - Lightning-Link®
返回頂端