Lightning-Link® HRP 抗體標定試劑盒

HRP Conjugation Kit - Lightning-Link®

貨號

ab102890

包裝

3 x 10 μg
100 μg
3 x 100 μg
1 mg
5 x 1 mg
5 mg

廠牌

Abcam

產品簡介

Abcam HRP Conjugation Kit - Lightning-Link® 提供了一種簡便快速的方法，可將 HRP 酵素（horseradish peroxidase，又稱為辣根過氧化物酶）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、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 HRP 酵素試劑小瓶，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 HRP 酵素試劑混合，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、ELISA、IHC 等實驗，無需額外處理。
Screen-printed carbon electrodes (SPCEs). Parkash, Om, et al used HRP Conjugation Kit - Lightning-Link® (ab102890) as part of the development and evaluation of a biosensor based on screen-printed carbon electrodes (SPCEs) for the detection of dengue-specific immunoglobulin M (IgM) antibodies. They used the kit to conjugate HRP to anti-dengue antibody for use in conjugation. (A) Schematic diagram of the screen printed carbon electrode (SCPE)-based dengue IgM biosensor. The biosensor was constructed by sequentially adding optimised concentration of anti-human IgM capture antibody, blocking agent, human IgM antibody or serum sample, dengue antigen and detection antibody, with washing steps in between. Electrochemical signal was generated following addition of TMB substrate. (B) Comparison of various immobilisation techniques for the goat anti-human IgM capture antibody. NC : negative control consisting of a dengue IgM negative serum sample; PC : dengue IgM positive serum sample. (C) Field Emission Scanning Electron Microscopy (FESEM) surface images of (left panel) a bare carbon electrode and (right panel) a carbon electrode modified with an anti-human IgM antibody using the streptavidin/biotin immobilisation system. Both capture and detection antibodies were labeled using Lightning-Link® conjugation kits (ab201796 and ab102890).
Screen-printed carbon electrodes (SPCEs). Parkash, Om, et al used HRP Conjugation Kit - Lightning-Link® (ab102890) as part of the development and evaluation of a biosensor based on screen-printed carbon electrodes (SPCEs) for the detection of dengue-specific immunoglobulin M (IgM) antibodies. They used the kit to conjugate HRP to anti-dengue antibody for use in conjugation. (A) Schematic diagram of the screen printed carbon electrode (SCPE)-based dengue IgM biosensor. The biosensor was constructed by sequentially adding optimised concentration of anti-human IgM capture antibody, blocking agent, human IgM antibody or serum sample, dengue antigen and detection antibody, with washing steps in between. Electrochemical signal was generated following addition of TMB substrate. (B) Comparison of various immobilisation techniques for the goat anti-human IgM capture antibody. NC : negative control consisting of a dengue IgM negative serum sample; PC : dengue IgM positive serum sample. (C) Field Emission Scanning Electron Microscopy (FESEM) surface images of (left panel) a bare carbon electrode and (right panel) a carbon electrode modified with an anti-human IgM antibody using the streptavidin/biotin immobilisation system. Both capture and detection antibodies were labeled using Lightning-Link® conjugation kits (ab201796 and ab102890). Image from Parkash, Om, et al., Diagnostics (Basel), 11(1):33; doi: 10.3390/diagnostics11010033. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Sandwich ELISA. HRP conjugation kit ab102890 is used by Abcam extensively in ELISA development. It is used for development / manufacturing of Abcam's SimpleStep ELISA® kits, including the Frataxin ELISA kit ab176112 used to produce the image above. Transformed B lymphocyte cells from Friedreich's Ataxia (FA) samples were compared to heterozygous carrier B lymphocyte cells (Carrier) and control B lymphocyte cells (Control). B lymphocyte cell extracts were analyzed across a 7-point titration (0.1-100 μg/mL) and frataxin levels were interpolated from the standard curve. Average interpolated values of Frataxin are plotted.
Sandwich ELISA. HRP conjugation kit ab102890 is used by Abcam extensively in ELISA development. It is used for development / manufacturing of Abcam's SimpleStep ELISA® kits, including the Frataxin ELISA kit ab176112 used to produce the image above. Transformed B lymphocyte cells from Friedreich's Ataxia (FA) samples were compared to heterozygous carrier B lymphocyte cells (Carrier) and control B lymphocyte cells (Control). B lymphocyte cell extracts were analyzed across a 7-point titration (0.1-100 μg/mL) and frataxin levels were interpolated from the standard curve. Average interpolated values of Frataxin are plotted.
ELISA. Cervin, Jakob, et al used HRP Conjugation Kit - Lightning-Link® (ab102890) as part of examining binding of Cholera Toxin subunit B (CTB) to blood group antigen LewisX (LeX). They used the kit to conjugate HRP to CTB and G33D (mutated version of CTB) for use in ELISA. ELISA with titrated amounts human serum albumin-linked oligosaccharides (HSA-OS), immobilized to wells and detected with (B) CTB-HRP and (C) G33D-HRP. Graph shows absorbance values from three independent experiments.
ELISA. Cervin, Jakob, et al used HRP Conjugation Kit - Lightning-Link® (ab102890) as part of examining binding of Cholera Toxin subunit B (CTB) to blood group antigen LewisX (LeX). They used the kit to conjugate HRP to CTB and G33D (mutated version of CTB) for use in ELISA. ELISA with titrated amounts human serum albumin-linked oligosaccharides (HSA-OS), immobilized to wells and detected with (B) CTB-HRP and (C) G33D-HRP. Graph shows absorbance values from three independent experiments. Image from Cervin, Jakob, et al., PLoS pathogens: 14.2 (2018): e1006862. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Competitive ELISA. Kristensen JH et al. used ab102890 for the quantification of neutrophil elastase (NE)-degraded elastin (EL). EL-NE fragment levels in serum from patients with IPF (n = 10) compared with controls (n = 9). ** p < 0.01. (B) : EL-NE fragment levels in serum from patients diagnosed with lung cancer (n = 40 in total), SCC (n = 16), adenocarcinoma (n = 16) and SCLC (n = 8) compared with controls (n = 12). **** p < 0.0001. Groups were compared by T-test with Welch correction. Data are shown as the geometric mean (95% CI). Abbreviations : IPF, idiopathic pulmonary fibrosis; SCC, squamous cell carcinoma; SCLC, small cell lung carcinoma.
Competitive ELISA. Kristensen JH et al. used ab102890 for the quantification of neutrophil elastase (NE)-degraded elastin (EL). EL-NE fragment levels in serum from patients with IPF (n = 10) compared with controls (n = 9). ** p < 0.01. (B) : EL-NE fragment levels in serum from patients diagnosed with lung cancer (n = 40 in total), SCC (n = 16), adenocarcinoma (n = 16) and SCLC (n = 8) compared with controls (n = 12). **** p < 0.0001. Groups were compared by T-test with Welch correction. Data are shown as the geometric mean (95% CI).
Abbreviations : IPF, idiopathic pulmonary fibrosis; SCC, squamous cell carcinoma; SCLC, small cell lung carcinoma. Image from Kristensen JH et al., BMC Pulm Med., 15:53. Fig 3.; doi: 10.1186/s12890-015-0048-5. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
ELISA. Gram M et al. used ab102890 to assess levels of cell free fetal haemoglobin (HbF) and haptoglobin (Hp) by ELISA. Samples were from normal pregnancies (Control) and women diagnosed with PE. The cell-free HbF plasma concentration of each patient sample (Control and PE) was plotted against the Hp plasma concentration.
ELISA. Gram M et al. used ab102890 to assess levels of cell free fetal haemoglobin (HbF) and haptoglobin (Hp) by ELISA. Samples were from normal pregnancies (Control) and women diagnosed with PE. The cell-free HbF plasma concentration of each patient sample (Control and PE) was plotted against the Hp plasma concentration. Image from Gram M et al.,PLoS One, 10(9):e0138111. Fig 1.; doi: 10.1371/journal.pone.0138111. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Western blot. Lemaire, Katleen, et al used HRP Conjugation Kit - Lightning-Link® (ab102890) as part of examining protein interactions. They used the kit to conjugate HRP to anti-BIP and anti-UFBP1 antibodies for use in detection by western blot after immunoprecipitation. Co-immunoprecipitation with BiP, UFL1 and UFM1 specific antibodies. BiP and UFBP1 antibodies were conjugated to HRP using ab102890 for detection after immunoprecipitation.
Western Blot. Lemaire, Katleen, et al used HRP Conjugation Kit - Lightning-Link® (ab102890) as part of examining protein interactions. They used the kit to conjugate HRP to anti-BIP and anti-UFBP1 antibodies for use in detection by western blot after immunoprecipitation. Co-immunoprecipitation with BiP, UFL1 and UFM1 specific antibodies. BiP and UFBP1 antibodies were conjugated to HRP using ab102890 for detection after immunoprecipitation. Image from Lemaire, Katleen, et al., PloS one, 6(4): e18517; doi: 10.1371/journal.pone.0018517. 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%)、磷酸鉀、氯化鈉、蔗糖、檸檬酸鈉、海藻糖等。疊氮化鈉 (Sodium Azide，≤0.1%) 雖然在緩衝液相容濃度範圍內不會干擾標定反應本身，但會不可逆地抑制 HRP 酵素活性。若樣本中含有疊氮化鈉，建議標定前先進行純化以去除疊氮化鈉，避免標定後的 HRP 接合物失去活性。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（> 4 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% BSA50% glycerol
0.1% sodium azide2PBSPotassium phosphate
Sodium chlorideHEPESSucrose
Sodium citrateEDTATrehalose
1 Tris buffered saline is almost always ≤ 50 mM / 0.6%
2 Sodium azide irreversibly inhibits HRP; antibodies with azide should be purified before using the HRP conjugation kit.
Incompatible buffer constituents
ThiomerosalProclinGlycine
ArginineGlutathioneDTT

套組內容物

Item3 x 10 µg100 µg3 x 100 µg1 mg5 x 1 mg5 mgStorage temperature
HRP mix3 vials1 vial3 vials1 vial5 vials1 vial-20°C
Modifier reagent1 vial1 vial1 vial1 vial1 vial1 vial-20°C
Quencher reagent1 vial1 vial1 vial1 vial1 vial1 vial-20°C

產品說明書

ab102890 HRP Conjugation Kit - Lightning-Link®
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