9-A5
CRL-1844 ™
- Product category
-
Animal cells
- Product type
-
Hybridoma
- Organism
-
Mus musculus (B cell); Mus musculus (myeloma), mouse (B cell); mouse (myeloma)
- Cell type
- hybridoma: b lymphocyte
- Morphology
- lymphoblast
- Applications
-
Immunology
- Product format
- Frozen
Documentation
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.
Detailed product information
General
- Specific applications
-
The antibody binds to and inhibits the catalytic activity of rat Na,K dependent ATPase.The antibody has been used to affinity purify the enzyme, and in immunohistocytochemical localization of the enzyme in rat tissues.
Characteristics
- Growth properties
- Suspension
- Derivation
-
Animals were immunized with purified rat Na,K dependent ATPase.Spleen cells were fused with P3/NSI/1-Ag4-1 myeloma cells.
- Genes expressed
- immunoglobulin, monoclonal antibody, against rat Na,K dependent ATPase
- Isotype
-
IgG1
- Comments
-
Animals were immunized with purified rat Na,K dependent ATPase.Spleen cells were fused with P3/NSI/1-Ag4-1 myeloma cells.The antibody binds to and inhibits the catalytic activity of rat Na,K dependent ATPase.The antibody appears to bind to the alpha subunit of the enzyme.The antibody has been used to affinity purify the enzyme, and in immunohistocytochemical localization of the enzyme in rat tissues.
Handling information
- Unpacking and storage instructions
-
- Check all containers for leakage or breakage.
- 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
-
The base medium for this cell line is ATCC Hybri-Care Medium, Catalog No. 46-X. Hybri-Care Medium is supplied as a powder and should be reconstituted in 1 L cell culture grade water. To make the complete growth medium, add the following components to the base medium:
- fetal bovine serum to a final concentration of 10%
- 1.5 g/L sodium bicarbonate for use with 5% CO2 in air atmosphere.
- Handling procedure
-
HANDLING PROCEDURE FOR FROZEN CELLS
- Initiate culture as soon as possible upon receipt.
- Thaw by rapid agitation in 37°C 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 37°C with 5% 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 replacement of
medium. Alternatively, cultures can be established by centrifugation with
subsequent resuspension at 10(5) viable cells/ml. Maintain cultures at a cell
concentration between 1 x 10(5) and 1 x 10(6) cells/ml. - Subculturing procedure
-
Medium Renewal: Every 2 to 3 daysCultures can be maintained by addition or replacement of fresh medium. Start cultures at 2 X 10 exp5 cells/ml and maintain between 1 X 10 exp5 and 1 X 10 exp6 cells/ml.
Quality control specifications
- Mycoplasma contamination
- Not detected
History
- Deposited as
- mouse (B cell); mouse (myeloma)
- Depositors
- HL Leffert
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.
Frequently Asked Questions
References
Curated Citations
Schenk DB, et al. Use of a monoclonal antibody to quantify (Na+, K+) ATPase activity and sites in normal and regenerating rat liver. J. Biol. Chem. 259: 14941-14951, 1984. PubMed: 6094583
Sutherland E, et al. Biochemical localization of hepatic surface-membrane Na+, K+-ATPase activity depends on membrane lipid fluidity. Proc. Natl. Acad. Sci. USA 85: 8673-8677, 1988. PubMed: 2847169
Schenk DB, Leffert HL. Monoclonal antibodies to rat Na+, K+-ATPase block enzymatic activity. Proc. Natl. Acad. Sci. USA 80: 5281-5285, 1983. PubMed: 6310568
Leffert HL, et al. Hepatic (Na+, K+)-ATPase: a current view of its structure, function and localization in rat liver as revealed by studies with monoclonal antibodies. Hepatology 5: 501-507, 1985. PubMed: 2987099
Hubert JJ, et al. Rat hepatic (Na+, K+)-ATPase: alpha-Subunit isolation by immunoaffinity chromatography and structural analysis by peptide mapping. Biochemistry 25: 4156-4163, 1986. PubMed: 3017414
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