Antigen Used for Immunization
A synthetic peptide of 20 amino acids (GIFMKPSELLKAEAHHAELC), which corresponded to residues 193–211 of human stromal cell-derived factor 2 (SDF-2).
Method of Immunization
The rat lymph node method established by Sado et al.(2)
Parental Cell Line Used for Fusion
Mouse myeloma SP2 cells.
Selection and Cloning Procedure
The hybridoma supernatants were screened using an enzyme linked immunosorbent assay (ELISA) against the human SDF-2 synthetic peptide. Positive clones were subcloned and rescreened by ELISA, immunoblotting, and immunoprecipitation.
Heavy and Light Chains of Immunoglobulin
Rat IgG2ακ.
Specificity
We determined specificity of MAb by immunoblotting analysis using the microsomes of SDF-2-knockout OCUM-2M/OXA cells. Our MAb gave specific signals as a 23 kDa band in the wild-type cells only.
Specific Antigen Identified
We did not identify specific antigen. However, it would include 20 amino acids (GIFMKPSELLKAEAHHAELC), which corresponded to residues 193–211 of human SDF-2.
Availability
| Yes | No | |
|---|---|---|
| Tissue culture supernatant | [ ] | [✓] |
| Ascitic fluid | [ ] | [✓] |
| Hybridoma cells | [✓] | [ ] |
References
- 1. Tanaka M, Shiota M, Koyama M, Nakayama J, Yashiro M, Semba K, and Goda N: Generation of rat monoclonal antibodies specific for human stromal cell-derived factor-2. Monolon Antib Immunodiagn Immunother 2020;39:23–26 [DOI] [PMC free article] [PubMed] [Google Scholar]
- 2. Sado Y, Kagawa M, Kishiro Y, Sugihara K, Naito I, Seyer JM, Sugimoto M, Oohashi T, and Ninomiya Y: Establishment by the rat lymph node method of epitope-defined monoclonal antibodies recognizing the six different alpha chains of human type IV collagen. Histochem Cell Biol 1995;104:267–275 [DOI] [PubMed] [Google Scholar]