标签：Vinculin 兔单克隆抗体

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

Primary Antibodies for Normalization

β-Actin Rabbit Monoclonal Antibody,β-肌动蛋白兔单克隆抗体

Primary Antibodies for Normalization

β-Actin Mouse Monoclonal Antibody,β-肌动蛋白小鼠单克隆抗体

Primary Antibodies for Normalization

COX IV Rabbit Primary Antibody,COX IV 兔一抗

Primary Antibodies for Normalization

GAPDH Rabbit Monoclonal Antibody,GAPDH 兔单克隆抗体

Primary Antibodies for Normalization

Histone H3 Mouse Monoclonal Antibody,组蛋白 H3 小鼠单克隆抗体

Primary Antibodies for Normalization

Histone H3 Rabbit Monoclonal Antibody

Primary Antibodies for Normalization

α-Tubulin Mouse Monoclonal Antibody

Primary Antibodies for Normalization

β-Tubulin Rabbit Polyclonal Antibody,β-微管蛋白兔多克隆抗体

Primary Antibodies for Normalization

Vinculin Rabbit Monoclonal Antibody,Vinculin 兔单克隆抗体

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.

“‘House-keeping’ proteins should not be used for normalization without evidence that experimental manipulations do not affect their expression.”

“Instructions for Authors” The Journal of Biological Chemistry. American Society for Biochemistry and Molecular Biology. Web. 3 March 2016.

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.

Primary Antibodies for Normalization Primary Antibodies for Normalization Primary Antibodies for Normalization

Western Blot Normalization and Publication Requirements

Sign up to review the science, best practices and mechanics of normalization strategies. Housekeeping proteins can be used as a strategy for Western blot normalization with the right experimental design. Find out what publishers need from you when using the housekeeping protein normalization strategy.

Lambda U® online learning portal is an online, on-demand resources can help ensure that you are getting the very best Western blot data.

注册以查看标准化策略的科学、最佳实践和机制。 通过正确的实验设计，管家蛋白可用作蛋白质印迹标准化的策略。 了解出版商在使用管家蛋白质标准化策略时需要您提供什么。

Lambda U® 在线学习门户是一个在线按需资源，可帮助确保您获得最佳的蛋白质印迹数据。

References

  1. Cazzalini O, Sommatis S, Tillhon M, Dutto I, Bachi A, Rapp A, Nardo T, Scovassi AI, Necchi D, Cardoso MC, Stivala LA, Prosperi E. (2014) CBP and p300 acetylate PCNA to link its degradation with nucleotide excision repair synthesis. Nucleic Acids Res. 42(13): 8433-48.
  2. Lanoix D, St-Pierre J, Lacasse A-A, Viau M, Lafond J, Vaillancourt C. (2012) Stability of reference proteins in human placenta: General protein stains are the benchmark. Placenta. 33: 151–156.
  3. Pérez-Pérez R, López JA, García-Santos E, Camafeita E, Gómez-Serrano M, Ortega-Delgado FJ, Ricart W, Fernández-Real JM, Peral B. (2012) Uncovering Suitable Reference Proteins for Expression Studies in Human Adipose Tissue with Relevance to Obesity. PLoS ONE 7(1): e30326.
  4. Barber RD, Harmer DW, Coleman RA, Clark BJ. (2005) GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiol Genomics. 21(3): 389-95.
  5. Rocha-Martins M, Njaine B, Silveira MS. (2012) Avoiding Pitfalls of Internal Controls: Validation of Reference Genes for Analysis by qRT-PCR and Western Blot throughout Rat Retinal Development. PLoS ONE 7(8): e43028.
  6. Eaton SL, Roche SL, Llavero Hurtado M, Oldknow KJ, Farquharson C, Gillingwater TH, Wishart TM. (2013) Total Protein Analysis as a Reliable Loading Control for Quantitative Fluorescent Western Blotting. PLoS ONE 8(8): e72457.
  7. Yu HR, Kuo HC, Huang HC, Huang LT, Tain YL, Chen CC, Liang CD, Sheen JM, Lin IC, Wu CC, Ou CY, Yang KD. (2011) Glyceraldehyde-3-phosphate dehydrogenase is a reliable internal control in western blot analysis of leukocyte subpopulations from children. Anal Biochem. 413(1): 24–9.
Primary Antibodies for Normalization Primary Antibodies for Normalization Primary Antibodies for Normalization

Western Blot Normalization and Publication Requirements

Sign up to review the science, best practices and mechanics of normalization strategies. Housekeeping proteins can be used as a strategy for Western blot normalization with the right experimental design. Find out what publishers need from you when using the housekeeping protein normalization strategy.

Lambda U® online learning portal is an online, on-demand resources can help ensure that you are getting the very best Western blot data.

LI-COR P/N 926-42215,Vinculin 兔单克隆抗体,LI-COR Vinculin 一抗

Reagents > Vinculin Rabbit Monoclonal Antibody for Normalization,用于标准化的 Vinculin 兔单克隆抗体

Vinculin Rabbit Monoclonal Antibody for Normalization

Vinculin is a highly conserved cytoskeletal protein. The Vinculin primary antibody can be used as an internal loading control for normalization and is particularly effective when detecting high molecular weight targets.

The expression of Vinculin, or any housekeeping protein (HKP), should be validated to ensure that its expression does not change under experimental conditions.

Once validated, Vinculin primary antibodies can be used for the detection of Vinculin when performing multiplex Western blot detection.

Detect Vinculin Rabbit Monoclonal Antibody with IRDye® Goat anti-Rabbit or IRDye Donkey anti-Rabbit secondary antibodies.

Other options for housekeeping protein normalization

Vinculin 是一种高度保守的细胞骨架蛋白。 Vinculin 一抗可用作标准化的内部上样对照，在检测高分子量目标时特别有效。

应验证 Vinculin 或任何管家蛋白 (HKP) 的表达，以确保其表达在实验条件下不会改变。

一旦通过验证，在进行多重蛋白质印迹检测时，Vinculin 一抗可用于检测 Vinculin。

使用 IRDye® Goat anti-Rabbit 或 IRDye Donkey anti-Rabbit 二抗检测 Vinculin Rabbit Monoclonal Antibody。

管家蛋白质标准化的其他选择

Reactivity and Specificity

Vinculin antibody is supplied in 10 mM HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and <0.02% sodium azide.

Do not aliquot the antibody.

Properties Vinculin Rabbit Monoclonal Antibody (P/N 926-42215)
Species Cross-Reactivity Human, mouse, rat, monkey
Target Molecular Weight 124 kDa
Isotype Rabbit IgG
Specificity/Sensitivity Detects endogenous levels of total vinculin protein and metavinculin, a 145 kDa splice variant of vinculin
Immunogen A recombinant protein that is specific to the amino terminus of human vinculin protein
Tested Application Western blot (WB), Immunohistochemistry (IHC), Flow Cytometry (F)

RRID

  • P/N 926-42215: RRID AB_2814925

Vinculin Rabbit Monoclonal Antibody Detected in C32 Cells

Vinculin Rabbit Monoclonal Antibody for Normalization
Vinculin Rabbit Monoclonal Antibody detected in C32 cells. C32 lysates were diluted from 2.5 μg to 156 ng. Lysates were separated on 4-12% Bis-Tris gels, electrophoresed at 200V for 45 minutes in MES Running Buffer, and transferred to nitrocellulose membranes in Tris-Glycine buffer at 100V for 65 minutes. Blots were blocked in Intercept® (PBS) Blocking Buffer (P/N 927-70001), probed with Vinculin Rabbit Monoclonal Antibody, and detected on an Odyssey® CLx Imager.
在 C32 细胞中检测到 Vinculin Rabbit Monoclonal Antibody。 C32 裂解物从 2.5 μg 稀释至 156 ng。 在 4-12% Bis-Tris 凝胶上分离裂解物，在 MES Running Buffer 中以 200V 电泳 45 分钟，然后在 100V 的 Tris-Glycine 缓冲液中转移至硝酸纤维素膜 65 分钟。 在 Intercept® (PBS) 封闭缓冲液（P/N 927-70001）中封闭印迹，用 Vinculin 兔单克隆抗体探测，并在 Odyssey® CLx Imager 上检测。