40 nm 奈米金粒子抗體標定試劑盒

Gold Conjugation Kit (40 nm, 20 OD)

貨號

ab154873

包裝

3 x 1 μg
10 x 1 μg
1 x 10 μg

廠牌

Abcam

產品簡介

傳統奈米金粒子標定方法多採用被動吸附 (passive adsorption) 方式，依賴抗體與裸露金屬表面之間的靜電作用力與疏水性交互作用吸附結合，這種方式須針對不同抗體逐一測試最適合的 pH 條件（通常需接近抗體的等電點），操作繁瑣且穩定性有限，抗體也可能因直接接觸金屬表面而發生變性，喪失親和力。Abcam Gold Conjugation Kit (40 nm, 20 OD) 採用的奈米金粒子表面則具備專利保護塗層，可完全隔絕抗體與金屬表面接觸，不易造成抗體結構破壞。標定反應是透過抗體上的離胺酸殘基 (lysine residues) 與奈米金粒子表面塗層形成共價鍵結，使抗體固定於奈米金粒子表面，標定效果不再受到抗體等電點影響，所有抗體皆可在同樣的 pH 條件、採用同一套標準流程完成標定，不僅操作簡易，所形成的標定產物也更為穩定。Abcam Gold Conjugation Kit 標定反應僅需將冷凍乾燥的奈米金粒子以待標定抗體溶液回溶即可啟動，動手操作時間僅約 2 分鐘，20 分鐘左右即可製備出 20 OD 的標定產物，全程無任何離心、沖洗或純化步驟，抗體回收率可達 100%。標定產物適用於側向流免疫分析法 (lateral flow assay, LFA) 快篩檢測平台。

ab154873 Gold Conjugation Kit (40 nm, 20 OD)
Lateral flow data. Antibody conjugation using the Gold Conjugation Kit (40 nm, 20 OD) (ab154873) vs traditional gold nanoparticle passive absorption techniques with uncoated gold nanoparticles, showing both enhanced signal intensity and improved specificity. 40 nm Gold particles were labeled with anti-IgA antibody and used to measure IgA concentration in a lateral flow inhibition assay, with IgA bound to a lateral flow strip.
Lateral flow assay. Vemulapati, S., et al used Gold Conjugation Kit (40nm, 20 OD) (ab154873) as part of examining vitamin D3 Deficiency. They used the kit to conjugate Gold to monoclonal anti-25(OH)D3 IgG antibody for use in lateral flow assay. Vitamin D3 Lateral Flow Assay: (A) Image and schematic of the 25(OH)D3 strip architecture and components. (B) Image and intensity plot of a participant with low Vitamin D3 and high T/C ratio (C) Image and intensity plot of a participant with healthy levels of 25(OH)D3 and low T/C ratio.
Lateral flow assay. Vemulapati, S., et al used Gold Conjugation Kit (40 nm, 20 OD) (ab154873) as part of examining vitamin D3 Deficiency. They used the kit to conjugate Gold to monoclonal anti-25(OH)D3 IgG antibody for use in lateral flow assay. Vitamin D3 Lateral Flow Assay: (A) Image and schematic of the 25(OH)D3 strip architecture and components. (B) Image and intensity plot of a participant with low Vitamin D3 and high T/C ratio (C) Image and intensity plot of a participant with healthy levels of 25(OH)D3 and low T/C ratio. Image from Vemulapati et al., Sci Rep., 7(1):14142; doi: 10.1038/s41598-017-13044-5. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Lateral flow assay. Lee, Seoho, et al used Gold Conjugation Kit (40 nm, 20 OD) (ab154873) as part of examining low-cost point-of-care quantification of vitamin B12 concentrations. They used the kit to conjugate Gold to anti-vitamin B12 IgG for use in competitive lateral flow assay. (A) Colorimetric variation of the test and control line regions on the silver enhanced B12 lateral flow test strip at different known concentrations of standard vitamin B12 samples (B) image processing algorithm used by the NutriPhone platform (C) Test and control line signals detected by the NutriPhone app as local intensity minima for 0 and 369 pmol/L of B12 standard samples (D) Calibration curve showing the T/C ratios of the colorimetric signals at different standard B12 concentrations. Image from Lee et al., Scientific reports 6 (2016): 28237. Sci Rep., 6:28237; doi: 10.1038/srep28237. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

主要特色

  • 採用共價鍵結技術，透過離胺酸殘基將抗體或蛋白質不可逆地固定於奈米金粒子表面的保護塗層上，標定產物比傳統被動吸附法製備的更加穩定。
  • 保護塗層可完全阻隔抗體與金屬表面直接接觸，因此標定效果與抗體的等電點無關，所有抗體皆可在同一個 pH 條件下完成標定，無需針對不同抗體逐一測試最適 pH 值。
  • 操作簡便快速，動手操作時間約 2 分鐘，20 分鐘左右即可完成標定，可達 100% 抗體回收率。
  • 奈米金粒子以冷凍乾燥形式提供，穩定性極佳，可承受極端條件（如在 70°C 以 2.5M NaOH 處理超過 1 小時仍可保持穩定），適用於多種物種來源的抗體（包括小鼠、兔、山羊、綿羊等），也適用於抗體片段的標定。

實驗注意事項

  • 待標定的抗體或蛋白質樣本建議溶於濃度較低（約 10 mM）的緩衝液中（如 MES、MOPS、HEPES），緩衝液 pH 值須介於 6.5–8.5；濃度過高的緩衝液可能會影響標定效果。不可使用 PBS 等含磷酸鹽的緩衝液；也應避免使用 BSA、Gelatin、Tris、Glycine、疊氮化鈉 (Sodium Azide)、Thimerosal (Thiomersal)、EDTA 或 Citrate 等螯合劑，也不可含有游離胺基或硫氫基成分（如胺基酸、乙醇胺、β-mercaptoethanol 或 DTT），這些成分皆可能干擾標定反應或影響奈米金粒子的穩定性。甘油 (glycerol) 濃度須低於 50%；糖類成分不影響標定效率，可正常使用。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。
  • 標定產物建議存放於 4°C，不建議存放於 -20°C；由於抗體與奈米金粒子之間形成的是共價鍵結，標定產物的整體穩定性主要取決於抗體本身的穩定性。
pH 6.5-8.5
Amine free buffer (e.g. MES, MOPS, HEPES) Yes
Sugars Yes
Glycerol < 50%
PBS No
Thiomersal No
Thimerosal No
Merthiolate No
Sodium Azide No
BSA No
Gelatin No
Tris No
Glycine No
Carboxylic acids (e.g. EDTA, Citrate) No
Nucleophilic components (Primary amines e.g. amino acids or ethanolamine and thiols e.g. mercaptoethanol or DTT) No

套組內容物

ItemQuantityStorage temperature
3 x 1 μg10 x 1 μg1 x 10 μg
Gold 40 nm3 vials10 vials1 vial-20°C
Gold 40 nm Reaction Buffer1 vial1 vial1 vial-20°C or +4°C
Gold 40 nm Antibody Diluent1 vial1 vial1 vial-20°C or +4°C
Gold 40 nm Quencher1 vial1 vial1 vial-20°C or +4°C

產品說明書

ab154873 Gold Conjugation Kit (40 nm, 20 OD)
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