Vinculin 兔单克隆抗体,β-肌动蛋白兔单克隆抗体用于标准化的一抗,Primary Antibodies for Normalization

When performing quantitative Western blots, internal loading controls and normalization are essential for reliable, precise comparison of protein expression. LI-COR offers several primary antibodies which can be used for two-color normalization when performing multiplex Western blots or for cell-based assay normalization, such as for In-Cell Western™ Assays.

在进行定量蛋白质印迹时，内部上样对照和标准化对于可靠、精确地比较蛋白质表达至关重要。 LI-COR 提供了多种一抗，可用于在进行多重蛋白质印迹或基于细胞的测定标准化（例如 In-Cell Western™ 测定）时进行双色标准化。

For Western blots, lysate preparation, sample loading, and membrane transfer introduce unavoidable variation. After validation, housekeeping proteins (HKP) can be used for normalization of protein levels.

• Normalize and correct for uneven loading, using the intensity of the internal loading control
• Visually compare protein levels with confidence, even if you don’t quantify bands
• Ensure that visually-observed changes in protein levels represent actual change, not artifacts

Read more about using a housekeeping protein as an internal loading control.

When using an HKP as your normalization strategy, it’s important to validate your HKP for each experiment to ensure its expression is stable. Many factors can influence expression including tissue, treatment, and cell density.

The Housekeeping Protein Validation Protocol can help you validate that your HKP expression is not changing with treatment. The Housekeeping Protein Normalization Protocol will guide you through the right experimental steps and calculations for normalization using an HKP. Validating your HKP is the only way to know that expression is not affected by the conditions of your experiment

管家蛋白质验证方案可以帮助您验证您的 HKP 表达不会随着治疗而改变。 管家蛋白质标准化方案将指导您完成正确的实验步骤和使用 HKP 进行标准化的计算。 验证您的 HKP 是了解表达式不受实验条件影响的唯一方法

For In-Cell Western Assays, a second protein target (such as actin, tubulin, COX IV, or GAPDH) can be used for normalization. Abundance of the normalization target must be unaffected by the cell treatments used.

Learn more about ICW assay normalization methods.

Comparison of Housekeeping Loading Controls

The table below has a few examples of some common housekeeping proteins, their molecular weights, and a few considerations. This list is not complete, and any housekeeping protein you choose should be fully validated for your system under study.