Revision 1

#9774Store at -20C

1 Kit

(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.

    使用に関する制限

    法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

    研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

    Revision 1
    #9774

    Loading Control Antibody Sampler Kit (Mouse)

    Loading Control Antibody Sampler Kit (Mouse): Image 1 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from Hela cells using β-Actin (8H10D10) Mouse mAb #3700. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    Loading Control Antibody Sampler Kit (Mouse): Image 2 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from COS-7 cells using α-Tubulin (DM1A) Mouse mAb #3873. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    Loading Control Antibody Sampler Kit (Mouse): Image 3 Expand Image
    Western blot analysis of extracts from various cell lines using Histone H3 (96C10) Mouse mAb.
    Loading Control Antibody Sampler Kit (Mouse): Image 4 Expand Image
    Western blot analysis of extracts from various cell types using β-Actin (8H10D10) Mouse mAb.
    Loading Control Antibody Sampler Kit (Mouse): Image 5 Expand Image
    Western blot analysis of extracts from HeLa and COS-7 cells using α-Tubulin (DM1A) Mouse mAb.
    Loading Control Antibody Sampler Kit (Mouse): Image 6 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.
    Loading Control Antibody Sampler Kit (Mouse): Image 7 Expand Image
    Western blot analysis of recombinant Actin isoforms using β-Actin (8H10D10) Mouse mAb (upper) and Pan-Actin Antibody #4968 (lower).
    Loading Control Antibody Sampler Kit (Mouse): Image 8 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using α-Tubulin (DM1A) Mouse mAb.
    Loading Control Antibody Sampler Kit (Mouse): Image 9 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using β-Actin (8H10D10) Mouse mAb.
    Loading Control Antibody Sampler Kit (Mouse): Image 10 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using α-Tubulin (DM1A) Mouse mAb (green). Red = Propidium Iodide (fluorescent DNA dye).
    Loading Control Antibody Sampler Kit (Mouse): Image 11 Expand Image
    Immunohistochemical analysis of paraffin-embedded human heart using β-Actin (8H10D10) Mouse mAb. Note the lack of staining of cardiac muscle.
    Loading Control Antibody Sampler Kit (Mouse): Image 12 Expand Image
    Flow cytometric analysis of HeLa cells using α-Tubulin (DM1A) Mouse mAb (solid line) compared to concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed line). Anti-mouse IgG (H+L), F(ab')2 fragment (Alexa Fluor 488 conjugate) #4408 was used as a secondary antibody.
    Loading Control Antibody Sampler Kit (Mouse): Image 13 Expand Image
    Confocal immunofluorescent analysis of NIH/3T3 cells using β-Actin (8H10D10) Mouse mAb (red) and PDI (C81H6) Rabbit mAb #3501 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Loading Control Antibody Sampler Kit (Mouse): Image 14 Expand Image
    Flow cytometric analysis of HeLa cells using β-Actin (8H10D10) Mouse mAb (solid line) compared to concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed line). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.