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28-6-20

CRL-2489

Product category
Animal cells
Product type
Hybridoma
Organism
Rattus norvegicus (B cell); Mus musculus (myeloma), rat (B cell); mouse (myeloma)
Cell type
hybridoma: lymphoblast b lymphocyte
Morphology
lymphoblast
Applications
Immunology
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Documentation

Biosafety Icon BSL 1

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.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Iscove's Modified Dulbecco's Medium (IMDM)

30-2005

Price: $24.00 ea

Fetal Bovine Serum (FBS)

30-2020

Price: $645.00 ea

Dimethylsulfoxide (DMSO)

4-X

Price: $49.00 ea

Detailed product information

General

Specific applications
The 28-2-7 (ATCC CRL-2487) and 28-6-20 (ATCC CRL-2489) antibodies bind the V kappa 24 light chain idiotypic specificity of MOPC167.
MOPC167 is an IgA myeloma specific for PC and HPCM27 is an IgM myeloma specific for PC.
The antibodies are specific for PC-binding proteins of VH1/V kappa 24 heavy and light chains of various isotypes.
These antibodies can be used as typing reagents for mouse IgA bearing the V kappa 24 idiotypic specificity.
They can be used in typing the 207-4 immunoglobulin transgenic mice that carry the VH1/V kappa 24 gene segment of the IgA PC specific MOPC167 myeloma.

Characteristics

Growth properties
Suspension
Derivation
Spleen cells were fused with P3X63Ag8.653 mouse myeloma cells.
Genes expressed
immunoglobulin, monoclonal antibody, against MOPC167 idiotype (V kappa 24)
Isotype
Rat IgG1
Rat IgG2a
Comments
The 28-2-7 (ATCC CRL-2487) and 28-6-20 (ATCC CRL-2489) antibodies bind the V kappa 24 light chain idiotypic specificity of MOPC167.
Animals were immunized with MOPC167 and HPCM27 anti-phosphocholine (PC) antibodies. MOPC167 is an IgA myeloma specific for PC and HPCM27 is an IgM myeloma specific for PC.
Spleen cells were fused with P3X63Ag8.653 mouse myeloma cells.
The antibodies are specific for PC-binding proteins of VH1/V kappa 24 heavy and light chains of various isotypes. They do not bind VH1/V kappa 22, VH1/V kappa 8, or VH1/V kappa 1 PC-binding proteins or other IgA or IgM myeloma proteins.
These antibodies can be used as typing reagents for mouse IgA bearing the V kappa 24 idiotypic specificity.
They can be used in typing the 207-4 immunoglobulin transgenic mice that carry the VH1/V kappa 24 gene segment of the IgA PC specific MOPC167 myeloma.

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete medium
Iscove's modified Dulbecco's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 85%; fetal bovine serum, 15%
Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure
To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at –70°C. Storage at –70°C will result in loss of viability.
  1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
  2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
  3. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete growth medium and spin at approximately 125 x g for 5 to 7 minutes. Discard supernatant.
  4. Resuspend the cell pellet with the recommended complete growth medium (see the specific batch information for the culture recommended dilution ratio) and dispense into a 25 cm2 or a 75 cm2 culture flask. It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
  5. Incubate the culture at 37°C in a suitable incubator. A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.
Subculturing procedure
Cultures can be maintained by addition of fresh medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 to 2  x 105 viable cells/mL.  Maintain cultures at a cell concentration between 5  x 104 and 1 x 106 cells/mL. Do not allow the cell concentration to exceed 1 x 106 cells/mL.

Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density)

Reagents for cryopreservation
Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X)

Quality control specifications

Mycoplasma contamination
Not detected

History

Deposited as
rat (B cell); mouse (myeloma)
Depositors
DG Sieckmann
Year of origin
1990

Legal disclaimers

Intended use
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.
Warranty

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.

Disclaimers

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.

Permits & Restrictions

Import Permit for the State of Hawaii

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.

This material was deposited by Donna G. Sieckmann, Naval Medical Research Center and is distributed for research purposes only under the ATCC Material Transfer Agreement. The Naval Medical Research Center has released the material subject to the following:

  1. Trasfers - the material or its products must not be transferred to third parties.
  2. Commercial Use - all for-profit and non-profit Recipients must obtain a commercial use license prior to Commercial Use.

Any proposed commercial use of these cells must first be negotiated with:

Naval Medical Research Center
The Commanding Officer
503 Robert Grant Avenue
Silver Spring, MD 20910-7500

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Curated Citations

Sieckmann DG, et al. Anti-idiotype monoclonal antibodies specific for the MOPC167 anti-phosphocholine transgene-encoded antibody. Hybridoma 16: 503-511, 1997. PubMed: 9455702

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.

Frequently Asked Questions

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Telephone

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