Store at -20°C
Chaps Cell
Extract Buffer
#9852
Supplied as a 10X stock
Orders n 877-616-CELL (2355)
[email protected]
Support n 877-678-TECH (8324)
[email protected]
Web n www.cellsignal.com
3 5 ml
n
(0.25 mL 1.25M DTT)
rev. 04/28/16
For Research Use Only. Not For Use In Diagnostic Procedures.
Description: Chaps Cell Extract Buffer can be used to
lyse cells under nondenaturing conditions and is recommended for the preparation of cytoplasmic cell lysates to be
used with our caspase signaling pathway antibodies (see
Companion Products).
Components:
10X CHAPS Cell Extract Buffer (5ml)
1X concentration:
50 mM Pipes/HCl (pH 6.5), 2 mM EDTA, 0.1% Chaps,
20 µg/ml Leupeptin, 10 µg/ml Pepstatin A,10 µg/ml
Aprotinin. Add DTT to 5mM.
1.25M (250X) DTT (0.25 mL)
1X concentration:
5 mM DTT
Preparation of 1X Cell Extract Buffer:
• Buffer should be prepared immediately before use.
• Immediately before dilution, invert the 10X Chaps
Cell Extract Buffer several times to suspend all buffer
components.
• Add 1/10 volume of 10X Chaps Cell Extract Buffer to 9/10
volume of Milli-Q water or equivalently purified water.
• Add DTT to final concentration of 5 mM (1:250 dilution)
and PMSF* to final concentration of 1 mM.
© 2014 Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
*reagent not included
Sample Preparation Using Chaps Cell
Extract Buffer:
(a) T reat cells by adding fresh media containing regulator
for desired time.
(b) Aspirate media from cultures; wash cells three times
with PBS; aspirate. Scrape cells into PBS and spin
down to pellet.
(c) L yse cells by adding ice cold 1X Chaps Cell Extract
Buffer (1 volume of cell pellet). Resuspend cells in the
buffer, freeze at -80°C and thaw twice, centrifuge the
lysate cold at 14,000 rpm. Keep the supernatant and
discard the pelleted cell debris.
(d) Add SDS Sample Buffer and heat sample to 95–100°C
for 5 minutes; cool on ice.
(e) M
icrocentrifuge for 5 minutes.
(f) L oad 5 µL onto SDS-PAGE gel (10 cm x 10 cm).
Storage: Store Chaps Cell Extract Buffer and 250X dithiothreitol
(DTT) at -20°C.
For application specific protocols please see the web
page for this product at www.cellsignal.com.
Please visit www.cellsignal.com for a complete listing
of recommended companion products.
Material Safety Data Sheet (MSDS) for
1 M DTT
New 03/08
I. IDENTIFICATION:
IX. EXPOSURE CONTROLS/PERSONAL PROTECTION:
Product name: 1 M DTT
Product Catalog Number: Component of kit #9002 and #9003
CAS number: None
Manufacturer Supplier: Cell Signaling Technology®
3 Trask Lane
Danvers, MA 01923 USA
1-978-867-2300 TEL
1-978-867-2400 FAX
1-978-578-6737 Emergency Phone
Appearance:
Odor:
pH:
Boiling Point:
Melting Point:
Freezing Point:
Volatile Organic Compounds (VOC):
Autoignition temp.
Solubility:
II. COMPOSITION/INFORMATION ON INGREDIENTS:
According to 29 CFR 1910.1200(d), hazardous ingredients at less than <1% and carcinogens
at less than < 0.1% therefore considered non-hazardous. Any hazardous or carcinogenic
ingredients meeting these criteria are listed below.
Ingredient
Dithiothreitol
CAS#
3483-12-3
Percent
15.4%
Irritant. Irritating to eyes, respiratory system, skin.
Target organs: Central nervous system.
Health: 2
Health: 2
Flammability: 0
Flammability: 0
Reactivity: 0
Reactivity: 0
Stability: Stable.
Conditions to avoid: Bases, oxidizing agents, reducing agents, alkali metals.
Hazardous Decomposition: Carbon monoxide, carbon dioxide, sulfur oxides, hydrogen
sulfide gas.
Hazardous polymerization: Will not occur.
Skin Exposure: Causes skin irritation. May be harmful if absorbed through skin.
Eye Exposure: Causes eye irritation.
Inhalation: May be harmful if inhaled. Material may be irritating to mucous membrane and
upper respiratory tract.
Ingestion: May be harmful if swallowed.
Signs and Symptoms of Exposure: Exposure can cause CNS depression.
IV. FIRST AID MEASURES:
XII. ECOLOGICAL INFORMATION:
Inhalation: Remove to fresh air. If breathing is difficult, get medical attention.
Ingestion: If person is conscious, wash out mouth with water. Get medical attention.
Skin Exposure: Wash skin with soap and water. If irritation develops or persists, get medical
attention.
Eye Exposure: Immediately flush eyes with water for at least 15 minutes. Get medical attention.
Data not available.
V. FIRE FIGHTING MEASURES:
XIV. TRANSPORT INFORMATION:
Flash Point: Not applicable.
Autoignition Temperature: Not applicable.
Explosion: Not applicable.
Fire Extinguishing Media: Water spray, dry chemical, alcohol foam, or carbon dioxide.
Firefighting: Wear protective clothing and self-contained breathing apparatus to prevent
contact with skin and eyes.
D.O.T.
Proper Shipping Name: None.
This substance is considered non-hazardous for transport.
IATA
Proper Shipping Name: None.
This substance is considered non-hazardous for air transport.
VI. ACCIDENTAL RELEASE MEASURES:
XV. REGULATORY INFORMATION:
Wear appropriate personal protective equipment as indicated in Section VIII. Absorb liquid with
an absorbent material. Transfer contaminated absorbent to a closed chemical waste container
for disposal. Wash spill site after material has been picked up for disposal.
EU Regulations/Classifications: Harmful. Harmful if swallowed. Irritating to eyes, respiratory system, skin.
VII. HANDLING AND STORAGE:
Store at -20°C in tightly closed container.
Do not breathe vapor. Avoid contact with eyes, skin, and clothing. Wash thoroughly after
handling.
VIII. EXPOSURE CONTROLS/PERSONAL
© 2008 Cell Signaling Technology, Inc.
X. STABILITY AND REACTIVITY:
XI. TOXICOLOGICAL INFORMATION:
III. HAZARD IDENTIFICATION:
HMIS Rating:
NFPA Rating:
Liquid
Unpleasant odor
Data not available
Data not available
Data not available
Data not available
Data not available
Data not available
Soluble in water
Ventilation System: A system of local (fume hood) and general exhaust is recommended.
Skin Protection: Wear compatible chemical resistant gloves and protective clothing.
Eye protection: Wear chemical safety goggles. Maintain emergency eyewash and shower in
work area.
Orders n 877-616-CELL (2355)
[email protected]
XIII. DISPOSAL CONSIDERATIONS:
Dispose of in accordance with federal, state and local environmental regulations. Contact a
licensed professional waste disposal service to dispose of this material.
US Regulations/Classifications: Toxic. Harmful if swallowed. Irritating to eyes, respiratory
system, skin. In case of contact with eyes, wash thoroughly and seek medical attention.
SARA Listed: No.
Canada (WHMIS): DSL: No, NDSL: No.
XVI. OTHER INFORMATION:
This product is not intended for use in humans. To the best of our knowledge, this document is
accurate. It is intended to serve as a guide for safe use of this product in a laboratory setting by
experienced personnel. The burden of safe use of this material rests entirely with the user. The
above information is believed to be accurate but is not necessarily all-inclusive and shall be
used only as a guide. Cell Signaling Technology, Inc., shall not be held liable for any damage
resulting from the handling of or from contact with the above product.
Support n 877-678-TECH (8324)
[email protected]
Web n www.cellsignal.com