測量蛋白質濃度(protein concentration)方法-BCA 蛋白定量

BCA蛋白定量分析套組 | VisualProtein貨號BC03-500  

BCA最常用的測量蛋白質濃度(protein concentration)的方法-Dual-Range BCA Protein Assay Kit

Dual-Range™ BCA 蛋白質檢測試劑盒(Dual-Range™ BCA Protein Assay Kit, BC03-500)是最常用的蛋白質檢測方法。與 Lowry 方法一樣，該測定依賴於蛋白質對 Cu2+ 離子的還原。通過與二辛可寧酸(bicinchoninate)反應轉化為紫色物質來檢測由此形成的 Cu+。 Dual-Range™ BCA Protein Assay Kit 與許多去污劑兼容，但與還原劑(reducing agents)如 DTT、DTE 和 2-巰基乙醇(2-Mercaptoethanol)等不兼容。二辛可寧酸 (bicinchoninic acid , BCA)，試劑 A 含有 BCA，試劑 B 則含有硫酸銅 (CuSO4)。 使用前混和兩種試劑，在鹼性環境下，蛋白質可將二價銅離 子 Cu2+ 還原為兩個單價銅離子 Cu+，而一個 Cu+ 可螯合兩 個 BCA 分子，使試劑顏色由蘋果綠轉變為紫色，此專一性呈 色與蛋白質濃度成正比，透過檢測 562nm 的吸光值，並比對 標準蛋白 ( 例如：BSA) 的校正曲線，可取得蛋白質濃度定量 數據。 用於 5–2,000 μg/mL 濃度範圍內的總蛋白質的比色檢測和定量。標準方案為 20-2000 μg/mL，增強方案為 5-250 μg/mL

● 寬線性範圍：標準方案為 20-2000 μg/mL，增強方案為 5-250 μg/mL

● BCA 對介面活性劑 (detergent) 具有高耐受性，不易受其影響

● 測定快速，可於 30 分鐘完成測定，比 Lowry 法快 4 倍

● 室溫保存，效期 24 個月

● 台灣生產，品質保證

圖 1.牛血清白蛋白 (BSA) 和牛丙種球蛋白 (BGG) 的標準曲線，採用微孔板程序（37°C，30 分鐘）。

圖 2.使用微孔板程序（37°C，30 分鐘），含 5% Triton X-100 或 8% SDS 的 BSA 標準曲線。

圖 3.使用微孔板程序和增強方案（60°C 30 分鐘）的微孔板程序的 BSA 標準曲線。

BC03-500    Dual-Range BCA Protein Assay Kit

> Reagent A  500 mL

> Reagent B  12 mL

BSA Standard (2 mg/mL) 1 mL x 10

 

Reference:

1. Young, Guang-Huar, et al. “The functional role of hemojuvelin in acute ischemic stroke.” Journal of Cerebral Blood Flow & Metabolism (2019): 0271678X19861448.

2. Chen, Sheng-Yi, et al. “Antioxidant and anti-inflammatory activities and bioactive compounds of the leaves of Trichodesma khasianum clarke.” Industrial Crops and Products 151 (2020): 112447.

3. Wang, Guan-Yu, et al. “Protective effect of rosmarinic acid-rich Trichodesma khasianum Clarke leaves against ethanol-induced gastric mucosal injury in vitro and in vivo.” Phytomedicine (2020): 153382.

4. Hsu, Jen‐Ying, et al. “Aqueous extract from Pepino (Solanum muricatum Ait.) leaves ameliorated insulin resistance, hyperlipidemia, and hyperglycemia in mice with metabolic syndrome.” Journal of Food Biochemistry 44.12 (2020): e13518.

5. Yang, Kai-Chiang, et al. “Effect of thermal treatments on the structural change and the hemostatic property of hair extracted proteins.” Colloids and Surfaces B: Biointerfaces 190 (2020): 110951.

6. Naito, Yoshisuke, et al. “Deterioration of Myofibrils Affected by Experimental Partial Immobilization in a Septic Rat Model.” Journal of St. Marianna University 11.2 (2020): 61-72.

7. Young, Guang‐Huar, et al. “Modulation of adenine phosphoribosyltransferase‐mediated salvage pathway to accelerate diabetic wound healing.” The FASEB Journal 35.3 (2021): e21296.

8. Leyton, Esteban, et al. “DEF8 and Autophagy-Associated Genes are Altered in Mild Cognitive Impairment, Probable Alzheimer’s Disease Patients and a Transgenic Model of the Disease.” Journal of Alzheimer’s Disease Preprint (2021): 1-16.

9. Hsu, Jen-Ying, et al. “Aqueous Extract of Pepino Leaves Ameliorates Palmitic Acid-Induced Hepatocellular Lipotoxicity via Inhibition of Endoplasmic Reticulum Stress and Apoptosis.” Antioxidants 10.6 (2021): 903.

10. Chen, Hong, et al. “Promoting effects of MiR-135b on human multiple myeloma cells via regulation of the Wnt/β-catenin/Versican signaling pathway.” Cytokine 142 (2021): 155495.

11. Chen, Ying-Yin, et al. “4‑Acetylantroquinonol B enhances cell death and inhibits autophagy by downregulating the PI3K/Akt/MDR1 pathway in gemcitabine‑resistant pancreatic cancer cells.” Oncology Letters 23.4 (2022): 1-13.

BR01-500	Bradford 蛋白質定量試劑	Dual-Range Bradford Reagent (1X), 500 ml ,
BR05-500	5X Bradford 蛋白質定量試劑	Dual-Range Bradford Reagent (5X), 500ml
BR05-500-K	5X Bradford 蛋白質定量試劑組	Dual-Range Bradford Protein Assay Kit, (500 ml +10* 2mg/ml BSA standard)
BP01-1L	西方墨點法遮蓋液	BlockPRO Blocking Buffer, 500mlx2
BF01-1L	西方墨點法遮蓋液-Protein Free	BlockPRO Protein-Free Blocking Buffer, 500mlx2
BF20-50P	西方墨點法遮蓋液-Protein Free	BlockPRO Protein-Free Blocking Buffer, 20X, 20包，每包2.5 mL， 使用前加入500mL ddH2O 回溶成 1X
		西方墨點法遮蓋液-Protein Free, 20X
BM01-500	西方墨點法遮蓋液-1分鐘配方	BlockPRO 1 Min Protein-Free Blocking Buffer, 500ml
SP01-500	抗體去除試劑	StripPRO 1 Min Stripping Buffer, 500ml
LF01-500	冷光試劑組- Prime	LumiFlash Prime Chemiluminescent Substrate,( 250mL solnA + 250mL soln B)
LF08-500	冷光試劑組- Ultima	LumiFlash Ultima Chemiluminescent Substrate, (250mL solnA + 250mL soln B)
LF16-500	冷光試劑組- Infinity	LumiFlash Infinity Chemiluminescent Substrate, (250mL solnA + 250mL soln B)
VC01-250	預染蛋白質標準液	VisColorTM Pre-Stained Protein Marker, 250ul x 1
VC01-500	預染蛋白質標準液	VisColorTM Pre-Stained Protein Marker, 250ul x 2
VC03-250	全範圍預染蛋白質標準液	VisColorTM Full-Range Pre-Stained Protein Marker, 250ul x 1
VC03-500	全範圍預染蛋白質標準液	VisColorTM Full-Range Pre-Stained Protein Marker, 250ul x 2
LH03-10	冷光筆	LuminolPen HRP system, (約 150 張 Western blot’s membranes  使用)
LH03-50	冷光筆	LuminolPen HRP system, (約1000 張 Western blot’s membranes  使用)
LH05-10	冷光筆 EZ版	LuminolPen EZ HRP System, (約 150 張 Western blot’s membranes 使用)
LH05-50	冷光筆 EZ版	LuminolPen EZ HRP System, (約 1000 張 Western blot’s membranes 使用)
RB10-1L	SDS-PAGE 電泳緩衝液	SDS-PAGE Running Buffer , 10X (Tris-Glycine, pH 8.3), 500 ml *2
RB500P	SDS-PAGE 電泳緩衝液濃縮包	SDS-PAGE Running Buffer , 10X (Tris-Glycine, pH 8.3), (20包，每 包須於使用前加入500mL ddH2O 回溶成 1X)
WTB10-1L	Western Blot Transfer Buffer	Western Blot Transfer Buffer, 10X (Tris-Glycine, pH 8.3), 500 ml *2  西方墨點法轉印緩衝液
WTB500P	西方墨點法轉印緩衝液濃縮包	Western Blot Transfer Buffer,(20包，每包須於使用前加入500mL ddH2O 回溶成 1X)
TBS10-1L	TBS緩衝液	TBS Buffer, 10X, 500 mL*2
TBS200P	TBS 緩衝液濃縮包	TBS Buffer, 25X, (30包，每包須於使用前加入200 mL ddH2O 回溶成 1X)
TBST10-1L	TBST 緩衝液	TBS Tween-20 Buffer, 10X, 500 mL*2
TBST200P	TBST 緩衝液濃縮包	TBS Tween-20 Buffer, 25X, (30包，每包須於使用前加入200 mL ddH2O 回溶成 1X)
PBS10-1L	PBS 緩衝液	PBS Buffer, 10X , 500 mL*2
PBS200P	PBS 緩衝液	PBS Buffer, 25X, (30包，每包須於使用前加入200 mL ddH2O 回溶成 1X)
PBST10-1L	PBST 緩衝液	PBS Tween-20 Buffer, 10X, 500 mL*2
PBST200P	PBST 緩衝液	PBS Tween-20 Buffer, 25X, (30包，每包須於使用前加入200 mL ddH2O 回溶成 1X)
SBN06-15	6x Laemmli SDS Sample Buffer, non-reducing	6x Laemmli SDS Sample Buffer, non-reducing, 15ml
SBR06-15	6x Laemmli SDS Sample Buffer, reducing	6x Laemmli SDS Sample Buffer, reducing, 15ml
VP01-125	全蛋白質負染色試劑組	VisPRO 5 Minutes Protein Stain Kit, (125mL soln 1 + 125mL soln2)
VP01-500	全蛋白質負染色試劑組	VisPRO 5 Minutes Protein Stain Kit, (500mL soln 1 + 500mL soln 2)
VP05-125	全蛋白質負染色試劑組- 五倍濃縮液	5x VisPRO 5 Minutes Protein Stain Kit, (125mL soln 1 + 125mL soln 2)
VP05-500	全蛋白質負染色試劑組- 五倍濃縮液	5x VisPRO 5 Minutes Protein Stain Kit, (250mL soln 1 + 250mL soln 2)
PP03-2C	磷酸化蛋質萃取試劑組	PhosPRO Phosphoprotein Enrichment Kit, (2 pre-packed column + 2mL Lysis buffer+250mL System buffer) +(100mL Elution buffer+100 mL Stringent buffer stock)
PP03-6C	磷酸化蛋質萃取試劑組	PhosPRO Phosphoprotein Enrichment Kit, (6 pre-packed column + 2mL Lysis buffer+250mL System buffer) + (100mL Elution buffer+100 mL Stringent buffer stock )
PP03-5E	磷酸化蛋質萃取試劑組	PhosPRO Phosphoprotein Enrichment Kit, (3 mL Resin + 2mL Lysis buffer+250mL System buffer) + (100mL Elution buffer+100 mL Stringent buffer stock)
PP05-2C		PhosPRO Phosphoprotein Purification Kit, (2 pre-packed column + 2mL Lysis buffer+250mL System buffer) +(100mL Elution buffer+100 mL Stringent buffer stock)
PP05-6C	磷酸化蛋白質純化試劑組	PhosPRO Phosphoprotein Purification Kit, (6 pre-packed column + 2mL Lysis buffer+250mL System buffer) + (100mL Elution buffer+100 mL Stringent buffer stock)  磷酸化蛋白質純化試劑組
PP05-5E	磷酸化蛋白質純化試劑組	PhosPRO Phosphoprotein Purification Kit, (3 mL Resin + 2mL Lysis buffer+250mL System buffer) + (100mL Elution buffer+100 mL Stringent buffer stock )
EP05-30	全蛋白質萃取試劑	ExtractPRO Protein Extraction Reagent
FP01-5	胞器蛋白質分離試劑組	FracPRO Protein Fractionation Kit, (10mL Soln1+ 10mL Soln2 +7.5mL Soln3+1.5mL Soln4)
FP01-50	胞器蛋白質分離試劑組	FracPRO Protein Fractionation Kit, (100mL Soln1 + 100mL Soln2+ 75mL Soln3 +15mL Soln4)
HB01-1L	促融合瘤細胞生長因子	HybriMore, Hybridoma cloning factor, for 1L medium dilution
IF01-4N	快速免疫佐劑	ImmunoFast Adjuvant, for 4 injections to mouse
DG03-1P	DIGE蛋白質標記試劑組	DIGE-Kit, (Cy2 400pmole + Cy3 400pmole + Cy5 400pmole+ 20ml DMF)
GLP01	負染膠片光箱	Gel Lighting Plate 21 x 29.7 cm(A4)
BLP01	正染膠片光箱	Band Lighting Plate 21 x 29.7 cm(A4)
BSB01	黑染盒	Black Staining Box 13 x 13 x 4 cm (For mini gel)
BSB02	黑染盒	Black Staining Box 17.5 x 17.5 x 4.5 cm (For regular gel)
BSB03	黑染盒	Black Staining Box 20 x 24 x 5 cm
BSB04	黑染盒	Black Staining Box 27 x 27 x 5 cm