Abbkine BCA蛋白定量試劑盒 | 貨號KTD3001 50-1000 µg/ml
Abbkine 蛋白定量試劑盒 (BCA 分析) 為總蛋白質的色度檢測和定量提供了一個簡單、快速、與洗滌劑相容的程式。該方法利用鹼性介質中蛋白質減少銅陽離子(Cu2+)到銅陽離子(Cu+)。生成的 Cu+ 與二面體酸 (BSA) 形成強烈的紫色複合物,在 562nm 時具有極強的吸收力。水溶性複合物的強度與樣品中的蛋白質量成正比。BCA蛋白測定適用於50-1000μg/ml範圍內的蛋白質濃度測量。
BCA (Bicinchoninic Acid , BCA) 在鹼性環境Cu+ 由鹼性Cu2+蛋白質的反應產生的。由此產生的反應和顏色是一種普通蛋白質定量方法的基礎,該方法能夠測量各種蛋白質濃度。增加蛋白質濃度會產生比例更深的顏色。BCA蛋白測定與勞里技術相比,BCA蛋白測定對普通干擾物質(如非離子洗滌劑和緩衝鹽)的耐受性更高。
Abbkine 蛋白定量試劑盒產品特點:
- 節省時間: 比傳統的Lowry method方法更容易,快四倍。
- 靈敏度: 檢測至 25 μg/ml,檢測蛋白的最小量達到5μg。
- 相容性: 不受大多數離子和非離子洗滌劑的影響。可與高達 5 % SDS、 5 % Triton X-100、 5 % Tween-20、 60、 80 在樣品中相容。
- 良好的線性度: 線性工作範圍從 50 到 1000 μg/ml.
- 與染料結合方法相比,蛋白質到蛋白質的變異性較少。
Fig. Abbkine Protein Quantification Kit (BCA Assay) standard curve
- Kit components
- BCA Reagent A (2x) • BCA Reagent B • BSA Standard
- Features & Benefits
- Time saving—much easier and four times faster than the classical Lowry method. • Sensitive—detect down to 25 µg/ml and the minimum amount of detection protein reaches 0.5μg. • Compatible—unaffected by most ionic and nonionic detergents. Can be compatible with up to 5 % SDS, 5 % Triton X-100, 5 % Tween-20, 60, 80 in the sample. • Good linearity—linear working range from 50 to 1000 µg/ml. • Excellent uniformity— exhibits less protein-to-protein variation than dye-binding methods.
- Calibration range
50-1000 µg/ml
- Limit of detection
25 µg/ml
- Usage notes
- The color of the reaction is not only related to the protein concentration of the sample, but also to the temperature of the reaction. If the concentration of protein in the sample is high (>50μg/ml), the reaction temperature is usually 37℃. If the concentration of protein in the sample is low (< 50μg/ml), the reaction temperature is 60℃. At this temperature, tryptophan, tyrosine and peptide bonds are fully oxidized, greatly improving the detection sensitivity.
• Commonly used concentrations of detergent SDS, Triton X-100, Tween do not affect the detection results, but results are affected by chelating agents (EDTA, EGTA), reducing agents (DTT, mercaptoethanol) and lipids. If such substance exists in the sample, try the Bradford method.
注意事項:
- 反應的顏色不僅與樣品的蛋白質濃度有關,而且與反應的溫度有關。如果樣品中的蛋白質濃度高(±50μg/ml),反應溫度通常為37°C。如果樣品中的蛋白質濃度低(=50μg/ml),反應溫度為60°C。在此溫度下,色氨酸、酪氨酸和肽鍵完全氧化,大大提高了檢測靈敏度。
- 常用的洗滌劑 SDS、Triton X-100、Tween 濃度不影響檢測結果,但結果受包紮劑(EDTA、EGTA)、還原劑(DTT、甲醇)和脂質影響。如果樣品中存在此類物質,請嘗試布拉德福德方法。
試劑4°C至少穩定12個月。 BCA試劑A和BCA 試劑B在4°C下穩定。BSA標準品自第一次融化後,應將BSA標準等分。 儲存在-20°C。
引用文獻
YL Zhang, C Geng, J Yang, J Fang, X Yan, PB Li. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2019.
技術提示
為避免交叉污染,添加試劑之間,添加樣品之間都需更換Tip。另外,每種試劑都應使用單獨的容器。BCA試劑B與BCA試劑A稀釋液混合後,觀察到渾濁混合後迅速消失,得到透明的綠色BCA工作溶液。BCA孵育時間和溫度都可以在相當廣泛的範圍內變化。使用更高的溫度和更長的孵育時間,蛋白質含量較低的樣品更容易定量。已知某些物質會干擾BCA分析,包括那些降低電位,螯合劑和強酸或強鹼。因為他們是已知即使在極低的濃度下也會干擾蛋白質估計,避免這些物質作為樣品緩衝液的成分。某些物質在較小程度上干擾了使用BCA進行蛋白質估算分析,並且在低於特定濃度的情況下,這些僅具有較小的(可忍受的)影響,如NP-40,SDS,Triton X-100,Tween-20等。在含有去污劑的溶液中測定蛋白質時,可通過以下方法獲得最佳結果向含有蛋白質標準品的孔中添加相同量的去污劑。
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