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ProCell Reagents

Patent Pending

ANTI-CLOTTING REAGENT PROTOCOL

CAT GGS-JL-00l

Pack size – 200 mg to perform 10 tests

PRODUCT DESCRIPTION

Anti-clotting Reagent is a newly developed product, consisting of a mixture of enzymes that actively break down the proteins that compose blood clots. Blood clots are hence quickly dissolved in precious cytogenetic samples, such as bone marrows and blood specimens that arrive clotted at the laboratory.

This product does not contain any streptokinase, which can be toxic to cells. Most clotted specimens that were previously lost due to clotting can now be rescued. Valuable bone marrow specimens arriving in containers with inadequate heparin can now be used to obtain metaphases, removing the requirement for repeat sampling in many cases. Follow Procedure 1 for bloods and bone marrows.

Anti-clotting Reagent is also effective on solid tumours, lymph node sections and CVS. These specimens often contain blood clots that hinder growth of the required cell type. Tests show that indirect CVS treated with Anti-Clotting Reagent are ready an average of 2 days earlier. Anti-clotting Reagent will also help to break down the tissues so colonies can be established more rapidly. Follow Procedure 2 for solid specimens.

STORAGE
Store the undissolved product at 4?C away from light for no more than 12 months. Frozen dissolved solution may be stored for up to 12 months. Never heat, freeze or expose the powdered product to drastic pH changes. For Laboratory research use only.

PROCEDURE 1

  1. Dissolve the sample of the Anticlotting Reagent in 100ml of PBS. Mix well by inverting the solution in a tube and filter sterilize using a 0.22 micron syringe filter once the powder is fully dissolved. This filtered solution should then be aliquoted into 10 ml lots and stored frozen until ready for use.
  2. Meanwhile centrifuge the clotted specimen in its original collection container for ten minutes.
  3. Replace the supernatant with 10ml of sterile Anticlotting Reagent solution and mechanically start to gently break the clots using a sterile glass pipette, for 30 seconds only.
  4. Incubate the specimen in a 37 degree Centigrade water bath until no clots can be seen.
  5. Most clots dissolve within five to ten minutes.
  6. When clots have dissolved, centrifuge the specimen for ten minutes.
  7. Discard supernatant and wash twice with culture media, centrifuging between each wash.
  8. Culture ‘buffy-coat’.

PROCEDURE 2

NOTE: This procedure should be used alongside a control

  1. Dissolve the sample of the Anticlotting Reagent in 100ml of PBS. Mix well by inverting the solution in a tube and filter sterilize using a 0.22 micron syringe filter once the powder is fully dissolved. This filtered solution should then be aliquoted into 5 ml lots and stored frozen until ready for use.
  2. Working with the sample in a Petri dish and making observations under an inverted microscope, add the 5 mls of Anticlotting reagent to the sample.
  3. Wait for 5 – 10 minutes, under the microscope the blood clots should be seen to dissolve and the surrounding solution should become red in colour.
  4. Pipette away all the debris and blood.
  5. Wash 2-3 times in PBS, pipetting away the solution each time.
  6. Set up the tissue in culture as normal.

QUALITY CONTROL

All batches are tested for ability to break down fibrin clots within 30 minutes.

MATERIAL SAFETY DATA SHEET

Date of issue 5th May 2005

IDENTIFICATION OF THE SUBSTANCE AND OF THE COMPANY

Catalogue Number: GGS-JL-00l Anti-clotting Reagent

For laboratory and research purposes only, removal and dispersion of blood clots from samples.

Genial Genetic Solutions Limited
The Heath Business and Technical Park, Runcorn, Cheshire WA7 4QX United Kingdom
Tel: +44 (0)870 757 9300
Fax: +44 (0)870 757 9301
Email: info@genialgenetics.com
Web: www.genialgenetics.com

COMPOSITION AND COMPONENT DATA (Working Solution)
Heparin and Enzymes in Dextrose/Saline solution

HAZARDS IDENTIFICATION
Anti-coagulant

FIRST AID MEASURES
In case of eye contact flush eyes with copious amounts of water. In case of skin contact immediately wash skin with soap and water.
If swallowed, wash out mouth with water and drink lots of fluids.
.
FIRE FIGHTING MEASURES
Not applicable.

ACCIDENTAL RELEASE MEASURES
Powder:-Collect by wiping with water dampened tissues.
Solution:-Mop up with tissues.
In both cases wear rubber gloves, and place used tissues in sealed plastic bag for proper disposal.

HANDLING AND STORAGE
Keep unused containers tightly closed. Store product at 4C for not more than expiry date

EXPOSURE CONTROL AND PERSONAL PROTECTION
Only work in a Class 2 biological safety cabinet.
Always wear a lab coat and rubber gloves.
Wash hands after handling.

PHYSICAL AND CHEMICAL PROPERTIES
Appearance: fine white powder
Boiling Point: As for water (prepared working solution)
Vapour Pressure (pascals): As for water (prepared working solution)
Specific Gravity: 1 (prepared working solution)
Flashpoint: N/A
Flammability Limits (%) Not applicable

STABILITY AND REACTIVITY
Stability: Stable as supplied
(see protocols for working solution stability)
Conditions to avoid: Not available
Incompatibility: Not available
Hazardous Decomposition By-products: Nil
Hazardous Polymerisation: Not available

TOXICOLOGICAL INFORMATION
To the best of our knowledge, the chemical, physical and toxicological properties have not been thoroughly investigated.
Not a known carcinogen.

ECOLOGICAL INFORMATION
Data not yet available

DISPOSAL CONSIDERATIONS
Use government approved contractor.
Observe all International, National, State and Local environmental laws.

TRANSPORT INFORMATION
Non-hazardous for transport

REGULATION
Not Applicable

OTHER INFORMATION
The above information is believed to be correct but does not purport to be all inclusive and shall be used only as a guide. Genial Genetic Solutions Ltd shall not be held liable for any damage resulting from handling or from contact with the above product.


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