M1/70.15.11.5.HL
TIB-128 ™
TIB-128 ™
ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
HANDLING PROCEDURE FOR FROZEN CELLS
- Initiate culture as soon as possible upon receipt.
- Thaw by rapid agitation in 37øC water bath. Thawing should be rapid (within
40-60 seconds). As soon as the ice is melted, remove the ampule from the water
bath and immerse in 70% ethanol at room temperature. All of the operations from
this point on should be carried out under strict aseptic conditions.
- The cells are supplied in two different types of glass ampules. One is a
standard ampule, the neck of which must be scored with a sharp file that has
been immersed in ethanol. A definitive sharp nick about 1/8" in length on one
side is necessary. The second type is prescored and is identifiable by a gold
band around the ampule neck, and should not be scored with a file.
- Break the neck of the ampule between several folds of a sterile towel.
- Transfer the cell suspension and dilute it with the recommended culture
medium in a culture flask (see specific batch information above for dilution
ratio); incubate at 37øC with 10% CO2 in air atmosphere. Since it is important
to avoid excessive alkalinity of the medium during recovery of the cells, it is
suggested that the culture medium be placed into the culture flask, tube, etc.
and the pH be adjusted, as necessary, prior to the addition of the ampule
contents. Note that the bicarbonate content of the culture medium will
determine whether an atmosphere containing CO2 will be required.
- It is not necessary to remove the freezing additive. However, if desired, the
culture medium may be changed to remove the protective freezing additive
(dimethylsulfoxide) 24 hours after thawing. If it is desired that the freezing
additive be removed immediately, or that a more concentrated cell suspension be
obtained, centrifuge the above diluted suspension at approximately 125 x g for
10 minutes, discard the fluid and resuspend the cells with growth medium at the
dilution ratio given in the specific batch information above.
FLUID RENEWAL
Add fresh medium (depending on cell density) every 2-3 days.
SUBCULTURE PROCEDURE
Cultures can be maintained by the addition of fresh medium or the replacement
of medium. Adherent cells can be dislodged by scraping and cultures established
by centrifugation with subsequent resuspension at 1-2 x 10(5) viable cells/ml.
Maintain cell culture density between 10(5) and 10(6) viable cells/ml.
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HANDLING PROCEDURE FOR FLASK CULTURES (SUSPENSION)
The flask was seeded with cells (see specific batch information above for
concentration), grown and completely filled with medium to prevent loss of
cells in transit. Upon receipt incubate the flask in an upright position for
several hours to return the flask contents to 37øC. After the temperature has
equilibrated, aseptically remove the entire contents of the flask and
centrifuge at 300 x g for 15 minutes. Resuspend the cell pellet in 10-12 ml of
the shipping medium. From this suspension remove a sample for a cell count and
viability so that the cell density of the suspension can be adjusted to 1-2 x
10(5) viable cells/ml. If the suspension needs to be diluted use the shipping
medium. Incubate the culture in a flat position at 37øC. The shipping medium
contains reduced sodium bicarbonate suitable for a 5% CO2 in air incubator.
DMEM usually contains 3.7 grams of sodium bicarbonate per liter and should be
incubated in a 10% CO2 in air incubator. Maintain the cell density of the
culture as suggested under the subculture procedure described above.
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NOTE
This material is available under the conditions that you will not use it for
commercial purposes or distribute it to third parties.
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CATALOGUE DESCRIPTION
This hybridoma secretes a monoclonal IgG2b antibody which reacts with the alpha
chain of the murine macrophage-granulocyte specific antigen Mac-1 (CD116).
Mac-1 is a macrophage differentiation antigen associated with type three
complement receptor. The antigen defined by this hybridoma is expressed in
large quantities on thioglycollate peritoneal exudate macrophages and in lesser
amounts on neutrophilic granulocytes, blood monocytes, 8% of spleen cells, 44%
of bone marrow cells, and less than 0.3% of thymus cells. This antibody
precipitates 2 polypeptides of 190,000 and 105,000 daltons. This antibody
labels human monocytes and polymorphonuclear leukocytes and a small population
of lymphocytes. It is capable of both natural killing (NK) and
antibody-dependent cellular cytotoxicity (ADCC). The hybridoma was formed by
the fusion of mouse myeloma line NS-1 with spleen cells from DA rats immunized
with C57BL/10 mouse spleen cells enriched for T lymphocytes. References: Eur.
J. Immunol. 8: 539-551, 1978; ibid., 9: 301-306, 1979; J. Biol. Chem. 256:
3833-3839, 1981; Monoclonal Antibodies, R. Kennett, et al. (eds.), pp. 185-217,
Plenum Press, 1980; J. Exp. Med. 158: 586, 1983. Originator: T. Springer,
Sidney Farber Cancer Institute, Harvard Medical School, Boston, MA.
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The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.
While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.
This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.
Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.
If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.
Springer T, et al. Monoclonal xenogeneic antibodies to murine cell surface antigens: identification of novel leukocyte differentiation antigens. Eur. J. Immunol. 8: 539-551, 1978. PubMed: 81133
Springer T, et al. Mac-1: a macrophage differentiation antigen identified by monoclonal antibody. Eur. J. Immunol. 9: 301-306, 1979. PubMed: 89034
Springer TA. Monoclonal antibody analysis of complex biological systems. Combination of cell hybridization and immunoadsorbents in a novel cascade procedure and its application to the macrophage cell surface. J. Biol. Chem. 256: 3833-3839, 1981. PubMed: 7217058
Sanchez-Madrid F, et al. Mapping of antigenic and functional epitopes on the alpha-and beta-subunits of two related mouse glycoproteins involved in cell interactions, LFA-1 and MAC-1. J. Exp. Med. 158: 586-602, 1983. PubMed: 6193226
Springer TACell -surface differentiation in the mouse: Characterization of Jumping and Lineage antigens using xenogeneic rat monoclonal antibodiesIn: Springer TAMonoclonal AntibodiesNew YorkPlenum Presspp. 185-217, 1980