Revision 1
#8980
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877-616-CELL (2355)
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3 Trask Lane | Danvers | Massachusetts | 01923 | USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Antibodies Included | Quantity | Application | Dilution |
---|---|---|---|
10 immunoprecipitations | ChIP | 1:50 | |
10 immunoprecipitations | ChIP | 1:50 | |
10 immunoprecipitations | ChIP | 1:50 |
Primers Included | Quantity | Application | Dilution |
---|---|---|---|
250 PCR reactions | ChIP | 1:10 | |
250 PCR reactions | ChIP | 1:10 |
Applications:
ChIP
Reactivity:
H
Sensitivity:
Endogenous
Product Usage Information
Directions for Use:A. Chromatin Immunoprecipitation:ChIP formulated antibodies have been tested and optimized using the SimpleChIP® Enzymatic Chromatin IP Kits (#9002 and #9003). Antibodies should be used at a dilution of 1:50 in a 500 μl ChIP reaction containing 10 to 15 μg of chromatin (4x106 cells). For the SimpleChIP® Enzymatic Chromatin IP protocol, please see the web page for this product at www.cellsignal.com.B. Quantification of DNA by qPCR:1. Label the appropriate number of PCR tubes or PCR plates compatible with the model of real-time PCR machine to be used. PCR reactions should be performed in duplicate and should include a tube with no DNA to control for contamination, and a serial dilution of a 2% total input chromatin DNA (undiluted, 1:5, 1:25, 1:125), which is used to create a standard curve and determine amplification efficiency.2. Add 2 μl of the appropriate ChIP DNA sample to each tube or well of the PCR plate.3. Prepare a master PCR reaction mix as described below. Add enough reagents for two extra reactions to account for loss of volume. Add 18 μl of the master PCR reaction mix to each PCR reaction tube or well of the PCR plate.Reagent Volume for 1 PCR Reaction (20 μl)
Nuclease-free H2O 6 μl
5 μM SimpleChIP® Primers 2 μl
2X SYBR-Green Reaction Mix 10 μl
4. Start the following PCR reaction program:
a. Initial Denaturation 95°C for 3 min
b. Denaturation 95°C for 15 sec
c. Anneal and Extension *Primer-specific temp. for 60 sec
d. Repeat steps b and c for a total of 40 cycles.*65 °C Anneal/Extension for SimpleChIP® Human Oct-4 Promoter Primers*60 °C Anneal/Extension for SimpleChIP® Human α Satellite Repeat Primers5. Analyze quantitative PCR results using software provided with the real-time PCR machine.
Storage
Primers are supplied in nuclease-free water at a concentration of 5 μM and should be stored at -20°C.
Specificity/Sensitivity
Source / Purification
Description
Background
Background References
- Looijenga, L.H. et al. (2003) Cancer Res 63, 2244-50.
- Boyer, L.A. et al. (2005) Cell 122, 947-56.
- Loh, Y.H. et al. (2006) Nat Genet 38, 431-40.
- Takahashi, K. and Yamanaka, S. (2006) Cell 126, 663-76.
- Okita, K. et al. (2007) Nature 448, 313-7.
- Yu, J. et al. (2007) Science 318, 1917-20.
- Okumura-Nakanishi, S. et al. (2005) J Biol Chem 280, 5307-17.
- Catena, R. et al. (2004) J Biol Chem 279, 41846-57.
- Rodda, D.J. et al. (2005) J Biol Chem 280, 24731-7.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Applications Key
ChIP: Chromatin IP
Cross-Reactivity Key
H: Human
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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.
Revision 1