Congo Red is an azo dye. Congo Red (CR) binding been used as a diagnostic test for the presence of amyloid in tissue sections.

Congo Red histochemical stain may serve as a simple screening tool for investigating if the aggregates in mutant cells have misfolded β-pleated sheet secondary structures. Congo Red histochemical dye has the ability to bind specifically to crossed β-pleated sheet structures. Wild-type HSPB1 should maintain protein homeostasis by binding proteins in non-native conformations, thereby preventing substrate aggregation. The T139M mutant, however, fails in this function and results in an accumulation of misfolded proteins, which are targeted by Congo Red for intercalating between the β-pleated sheet structures. Congo Red histochemical stain may serve as a simple tool to investigate if the aggregates in mutant cells have misfolded β-pleated sheet secondary structures^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

696.66

C₃₂H₂₂N₆Na₂O₆S₂

573-58-0

刚果红

Room temperature in continental US; may vary elsewhere.

4deg;C, sealed storage, away from moisture and light

*In solvent : -80deg;C, 6 months; -20deg;C, 1 month (sealed storage, away from moisture and light)

DMSO : 20 mg/mL (28.71 mM; Need ultrasonic)

H₂O : < 0.1 mg/mL (insoluble)

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂：;10% DMSO ;; 40% PEG300 ;; 5% Tween-80 ;; 45% saline

  Solubility: ≥ 2 mg/mL (2.87 mM); Clear solution

  此方案可获得 ≥ 2 mg/mL (2.87 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂：;10% DMSO ;; 90% (20% SBE-β-CD in saline)

  Solubility: ≥ 2 mg/mL (2.87 mM); Clear solution

  此方案可获得 ≥ 2 mg/mL (2.87 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.0 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明

• [1]. Amornvit J, et al. A novel p.T139M mutation in HSPB1 highlighting the phenotypic spectrum in a family. Brain Behav. 2017 Jul 21;7(8):e00774.

HeLa cells are selected for these studies due to their large cytoplasmic volume. Cells transfected with mutant or wild-type HSPB1 constructs are grown on coverslips for 24 hr and then are stained with Congo red to determine if the aggregates display amyloidogenic properties. Briefly, cells are first fixed with 10% formalin for 10 min and stained with 1% Congo red for 5 min, followed by destaining with 0.01% potassium hydroxide in 50% ethanol. Coverslips are then passed through graded ethanol concentrations for dehydration and mounted in a mounting medium and examined by fluorescent microscopy under rhodamine filter^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Amornvit J, et al. A novel p.T139M mutation in HSPB1 highlighting the phenotypic spectrum in a family. Brain Behav. 2017 Jul 21;7(8):e00774.