Revision 1

#9774Store at -20C

1 个试剂盒

(3 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
β-Actin (8H10D10) Mouse mAb 3700 20 µl 45 kDa Mouse IgG2b
α-Tubulin (DM1A) Mouse mAb 3873 20 µl 52 kDa Mouse IgG1
Histone H3 (96C10) Mouse mAb 3638 20 µl 17 kDa Mouse IgG1
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Loading Control Antibody Sampler Kit provides an economical means to detect a variety of housekeeping proteins. The kit contains enough primary and secondary antibodies to perform two western blot experiments.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

Housekeeping proteins perform numerous basic functions within the cell and are constitutively expressed at high levels in a variety of tissues and cell types. Western blot analysis commonly uses housekeeping proteins such as β-actin, histone H3, and α-tubulin as loading controls. Actin is a ubiquitous protein and a major component of the eukaryotic cytoskeleton. Actin exists mainly as the F-actin fibrous polymer (1). Globular tubulin subunits made up of α- and β-tubulin heterodimers are the building blocks of microtubules, one of three types of cytosolic fibers that comprise the cytoskeleton (2). Histone proteins, including histone H3, make up the primary building block of chromatin known as nucleosomes. Modulation of the chromatin structure plays an important role in the regulation of transcription in eukaryotes (3).

  1. Condeelis, J. (2001) Trends Cell Biol 11, 288-93.
  2. Westermann, S. and Weber, K. (2003) Nat Rev Mol Cell Biol 4, 938-47.
  3. Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    限制使用

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    Revision 1
    #9774

    Loading Control Antibody Sampler Kit (Mouse)

    Loading Control Antibody Sampler Kit (Mouse): Image 1 Expand Image
    使用 β-Actin (8H10D10) Mouse mAb #3700 对 Hela 细胞的裂解物 (0.1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的单一靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
    Loading Control Antibody Sampler Kit (Mouse): Image 2 Expand Image
    使用 α-Tubulin (DM1A) Mouse mAb #3873 对 COS-7 细胞的裂解物 (0.1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
    Loading Control Antibody Sampler Kit (Mouse): Image 3 Expand Image
    使用 Histone H3 (96C10) Mouse mAb 对来自不同细胞系的提取物进行蛋白质印迹分析。
    Loading Control Antibody Sampler Kit (Mouse): Image 4 Expand Image
    使用 β-Actin (8H10D10) Mouse mAb 对来自不同细胞类型的提取物进行蛋白质印迹分析。
    Loading Control Antibody Sampler Kit (Mouse): Image 5 Expand Image
    使用 α-Tubulin (DM1A) Mouse mAb 对来自 HeLa 细胞和 COS-7 细胞的提取物进行蛋白质印迹分析。
    Loading Control Antibody Sampler Kit (Mouse): Image 6 Expand Image
    一抗与靶标蛋白结合之后,与偶联 HRP 的二抗形成复合体。添加 LumiGLO*,它可在酶催化的分解期间发光。
    Loading Control Antibody Sampler Kit (Mouse): Image 7 Expand Image
    使用 β-Actin (8H10D10) Mouse mAb(上图)和 Pan-Actin Antibody #4968(下图)对重组肌动蛋白同工型进行蛋白质印迹法分析。
    Loading Control Antibody Sampler Kit (Mouse): Image 8 Expand Image
    使用 α-Tubulin (DM1A) Mouse mAb 对石蜡包埋的人结肠癌进行免疫组织化学分析。
    Loading Control Antibody Sampler Kit (Mouse): Image 9 Expand Image
    使用 β-Actin (8H10D10) Mouse mAb 对石蜡包埋的人乳腺癌进行免疫组织化学分析。
    Loading Control Antibody Sampler Kit (Mouse): Image 10 Expand Image
    使用 α-Tubulin (DM1A) Mouse mAb(绿)对 Hela 细胞进行共聚焦免疫荧光分析。红色 = 碘化丙啶(DNA 荧光染料)。
    Loading Control Antibody Sampler Kit (Mouse): Image 11 Expand Image
    使用 β-Actin (8H10D10) Mouse mAb 对石蜡包埋的人心脏进行免疫组织化学分析。注意缺少心肌染色。
    Loading Control Antibody Sampler Kit (Mouse): Image 12 Expand Image
    以浓度匹配的 Mouse (G3A1) mAb IgG1 Isotype Control #5415(虚线)作为对照,使用 α-Tubulin (DM1A) Mouse mAb(实线)对 HeLa 细胞进行流式细胞分析。Anti-mouse IgG (H+L), F(ab')2 fragment (Alexa Fluor 488 conjugate) #4408 用作二抗。
    Loading Control Antibody Sampler Kit (Mouse): Image 13 Expand Image
    使用 β-Actin (8H10D10) Mouse mAb (红)和 PDI (C81H6) Rabbit mAb #3501(绿)对 NIH/3T3 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
    Loading Control Antibody Sampler Kit (Mouse): Image 14 Expand Image
    以浓度匹配的 Mouse (G3A1) mAb IgG1 Isotype Control #5415(虚线)作为对照,使用 β-Actin (8H10D10) Mouse mAb(实线)对 HeLa 细胞进行流式细胞分析。Anti-mouse IgG (H+L),F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 作为二抗。