标签：用于 IRDye® 二抗

硝酸纤维素膜的 NewBlot™ Nitro 剥离缓冲液,用于 IRDye® 二抗,IRDye 800CW,IRDye 680RD, IRDye 680LT

Reagents > NewBlot™ Nitro Stripping Buffer for Nitrocellulose Membranes,用于硝酸纤维素膜的 NewBlot™ Nitro 剥离缓冲液

NewBlot™ Nitro Stripping Buffer for Nitrocellulose Membranes

NewBlot Nitro is optimized for near-infrared (NIR) Western blots and is specially formulated for use with IRDye® secondary antibodies, including IRDye 800CW, IRDye 680RD, and IRDye 680LT.

NewBlot Nitro is intended for nitrocellulose membranes only, and is ideal for removal of secondary antibody.

NewBlot Nitro 针对近红外 (NIR) 蛋白质印迹进行了优化，专门配制用于 IRDye® 二抗，包括 IRDye 800CW、IRDye 680RD 和 IRDye 680LT。

NewBlot Nitro 仅适用于硝酸纤维素膜，是去除二抗的理想选择。

Choosing the Right Stripping Buffer

Specifications

NewBlot Nitro Stripping Buffer is supplied as a 5X concentrated solution. Each 100 mL bottle is sufficient for up to 3000 cm2 or approximately fifty 7 x 8.5 cm Odyssey® nitrocellulose membranes.

NewBlot Nitro stripping buffer contains hazardous materials and additional shipping costs will be applied.

NewBlot Nitro Stripping Buffer 以 5X 浓缩溶液的形式提供。 每个 100 mL 瓶足以容纳 3000 cm2 或大约 50 个 7 x 8.5 cm Odyssey® 硝酸纤维素膜。

NewBlot Nitro 剥离缓冲液含有有害物质，将收取额外的运输费用。

Example Data

NewBlot™ Nitro Stripping Buffer for Nitrocellulose Membranes
Example of stripping and reprobing with NewBlot Nitro Stripping Buffer. Rabbit anti-β-tubulin and mouse anti-ERK2 were run on a nitrocellulose membrane Western blot. The blot was probed with IRDye 680 Goat anti-Rabbit (red) and IRDye 800CW Goat anti-Mouse (green). The blot was stripped with NewBlot Nitro Stripping Buffer (1X) and reprobed three sequential times with the same antibodies. All blots have been scanned at 169 micron resolution and scan intensity of 6 on the Odyssey Infrared Imaging System.

Factors Affecting Efficiency when Using NewBlot Nitro Stripping Buffer

Recommended Standard Conditions: 1X NewBlot Nitro, 5 minutes at room temperature.

Below are key factors that affect stripping efficiency with NewBlot Nitro on Odyssey nitrocellulose membranes. The data figure above compares stripping efficiency using various concentrations, time, and temperature to the recommended standard conditions, 1X, 5 min, ambient.

For optimal stripping results, follow the optimization guidelines in NewBlot Nitro Stripping Buffer pack insert.

以下是影响 Odyssey 硝酸纤维素膜上使用 NewBlot Nitro 剥离效率的关键因素。 上面的数据图比较了使用各种浓度、时间和温度与推荐的标准条件（1X，5 分钟，环境）的剥离效率。

为获得最佳剥离结果，请遵循 NewBlot Nitro Stripping Buffer pack insert 中的优化指南。

Amount of time blot is in stripping buffer

Increasing stripping time has the greatest effect on efficiency.

Increasing the stripping time may lead to increased damage/loss of target antigens, and reduce the success of reprobing.

Sample type and preparation

Even under the most stringent stripping conditions, the fluorescent signal may not be removed completely due to sample load amount, antibody affinity/avidity, and target protein abundance.

即使在最严格的剥离条件下，由于样品上样量、抗体亲和力/亲合力和靶蛋白丰度，荧光信号也可能无法完全去除。

Blot handling conditions

Washing, scanning, or stripping efficiency will be affected if the blot is allowed to dry at all during incubation. Keep the blot moist at all times.

Buffer concentration and temperature used for stripping

Increasing the stripping buffer concentration and temperature significantly improves stripping effectiveness but can also have a highly detrimental effect on reprobing.

If the optimization process given in the protocol does not produce the desired stripping results, stripping buffer incubation can be carried out at 37 °C using a water bath or warm-air incubator.

Do not microwave the NewBlot Nitro Stripping Buffer or the nitrocellulose blot.

增加剥离缓冲液浓度和温度会显着提高剥离效果，但也会对重新探测产生非常不利的影响。

如果协议中给出的优化过程没有产生所需的剥离结果，则可以使用水浴或暖空气培养箱在 37°C 下进行剥离缓冲液孵育。

不要微波 NewBlot Nitro 剥离缓冲液或硝酸纤维素印迹。