Revision 1

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
beta-Actin (8H10D10) Mouse Monoclonal Antibody370020 µl45 kDaMouse IgG2b
alpha-Tubulin (DM1A) Mouse Monoclonal Antibody387320 µl52 kDaMouse IgG1
Histone H3 (96C10) Mouse Monoclonal Antibody363820 µl17 kDaMouse IgG1
Anti-mouse IgG, HRP-linked Antibody7076100 µlHorse

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).

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.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using Histone H3 (96C10) Mouse mAb.
undefined Image 1: Loading Control Antibody Sampler Kit (Mouse)
Western blot analysis of extracts from various cell types using β-Actin (8H10D10) Mouse mAb.
undefined Image 2: Loading Control Antibody Sampler Kit (Mouse)
Western blot analysis of extracts from HeLa and COS-7 cells using α-Tubulin (DM1A) Mouse mAb.
undefined Image 3: Loading Control Antibody Sampler Kit (Mouse)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
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.
undefined Image 4: Loading Control Antibody Sampler Kit (Mouse)
Western blot analysis of recombinant Actin isoforms using β-Actin (8H10D10) Mouse mAb (upper) and Pan-Actin Antibody #4968 (lower).
undefined Image 5: Loading Control Antibody Sampler Kit (Mouse)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using α-Tubulin (DM1A) Mouse mAb.
undefined Image 6: Loading Control Antibody Sampler Kit (Mouse)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using β-Actin (8H10D10) Mouse mAb.
undefined Image 7: Loading Control Antibody Sampler Kit (Mouse)
Confocal immunofluorescent analysis of HeLa cells using α-Tubulin (DM1A) Mouse mAb (green). Red = Propidium Iodide (fluorescent DNA dye).
undefined Image 8: Loading Control Antibody Sampler Kit (Mouse)
Immunohistochemical analysis of paraffin-embedded human heart using β-Actin (8H10D10) Mouse mAb. Note the lack of staining of cardiac muscle.
undefined Image 9: Loading Control Antibody Sampler Kit (Mouse)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
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.
undefined Image 10: Loading Control Antibody Sampler Kit (Mouse)
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).
undefined Image 11: Loading Control Antibody Sampler Kit (Mouse)
Flow cytometric analysis of HeLa cells using β-Actin (8H10D10) Mouse mAb (solid line) compared to concentration-matched Mouse (E7Q5L) mAb IgG2b Isotype Control #53484 (dashed line). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
undefined Image 12: Loading Control Antibody Sampler Kit (Mouse)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Simple WesternTM analysis of lysates (1.0 mg/mL) from HeLa cells using Histone H3 (96C10) Mouse mAb #3638. The virtual lane view (left) shows the target band (as indicated) at 1:50 dilution of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 dilution of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 2-40 kDa separation module.
undefined Image 13: Loading Control Antibody Sampler Kit (Mouse)
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.
undefined Image 14: Loading Control Antibody Sampler Kit (Mouse)
Western blot analysis of extracts from HeLa cells (lane 1) and C2C12 cells (lane 2), using beta-Actin (8H10D10) Mouse Monoclonal Antibody #3700. Anti-mouse IgG (H+L) (DyLight 680 Conjugate) #5470 (red) (Panel A) or Anti-mouse IgG (H+L) (DyLight 800 4X PEG Conjugate) #5257 (Panel B) were used as secondary antibodies.
undefined Image 15: Loading Control Antibody Sampler Kit (Mouse)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
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.
undefined Image 16: Loading Control Antibody Sampler Kit (Mouse)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.