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35.1
Tissue
Spleen (B cell origin) / Bone marrow (NSI/1 myeloma fusion partner)
Disease
Hybridoma (B cell × myeloma fusion); produces monoclonal antibody against human CD2 (Tp50)
Metastatic site
Not applicable (hybridoma cell line; not a primary tumor sample)
Synonyms
HB-222

Documents

35.1

Cell line · Product number 305002

35.1 are hybridoma cells derived from a Mus musculus B cell and myeloma. Of particular interest is their reactivity to the human T cell surface protein Tp50 ...

€700.00excl. tax and shipping

Customization / Recommendations

In stock
Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description
35.1 are hybridoma cells derived from a Mus musculus B cell and myeloma. Of particular interest is their reactivity to the human T cell surface protein Tp50 (CD2), making them valuable tools for targeted research. The antibody produced by 35.1 cells inhibits T-cell proliferative responses and cytotoxicity without interfering with antibody-mediated cellular cytotoxicity. Derived from the fusion of spleen cells with NSI/1 myeloma cells, 35.1 cells express genes for immunoglobulin production, specifically monoclonal antibodies against human CD2. The isotype of the antibody is IgG2a.
Tissue
Spleen (B cell origin) / Bone marrow (NSI/1 myeloma fusion partner)
Disease
Hybridoma (B cell × myeloma fusion); produces monoclonal antibody against human CD2 (Tp50)
Metastatic site
Not applicable (hybridoma cell line; not a primary tumor sample)
Applications
Monoclonal antibody production (anti-human CD2, IgG2a); T cell biology research; CD2-mediated T cell activation and proliferation studies; immunological assays requiring CD2 blockade; cytotoxicity research; hybridoma technology
Synonyms
  • HB-222

02Classification

Organism
Mouse
Sex
Unspecified
Growth properties
Suspension
GMO status
GMO-S1
GMO status details
GMO-S1: This hybridoma contains rearranged immunoglobulin genes from a B cell-myeloma fusion. IgG2a heavy and light chain genes are expressed from the B cell fusion partner. This classification applies only within Germany and may differ elsewhere.
Biosafety level
2

03Characteristics

Morphology
Lymphoblast
Cell type
B lymphoblast / hybridoma cells
Doubling time
approx. 24 to 36 hours

04Identifiers

Citation
1
NCBI TaxID
10090
Cellosaurus
CVCL_7239

05Expression and susceptibility

Protein expression
Immunoglobulin, monoclonal antibody, against human CD2

06Handling and culture

Culture medium
RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
Incubation atmosphere

37°C, 5% CO2, humidified atmosphere.

Supplements
Supplement the medium with 10% FBS, 0,05 mM 2-mercaptoethanol
Dissociation reagent
None
Subculturing
Gently homogenize the cell suspension in the flask by pipetting up and down, then take a representative sample to determine the cell density per ml. Dilute the suspension to achieve a cell concentration of 1 x 105 cells/ml with fresh culture medium, and aliquot the adjusted suspension into new flasks for further cultivation.
Split ratio
1 to 5
Seeding density
1 to 5 × 10⁵ cells/ml
Fluid renewal
2 to 3 times per week
Post-thaw recovery
After thawing, centrifuge the cells at 300×g for 5 minutes, remove the cryoprotectant-containing supernatant, and resuspend in fresh pre-warmed complete medium at 1 × 10⁵ cells/ml. Allow at least 24–48 hours for recovery and expansion before the first passaging step.
Thawing and culturing

  1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

  2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

  3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

  4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

  5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

  6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

07Quality control and cryopreservation

Sterility

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

Freeze medium
As a cryopreservation medium, we use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
Storage conditions

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Shipping conditions

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

08Donor

Breed / subspecies
BALB/c (NSI/1 myeloma background)
Donor age
Age unspecified
Ethnicity
Not applicable (mouse cell line)