Monoclonal Antibody Against Stromal Cell-Derived Factor 2.

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.(

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.

	Yes	No
Tissue culture supernatant	[ ]	[✓]
Ascitic fluid	[ ]	[✓]
Hybridoma cells	[✓]	[ ]