Fig. 1.

Specificity of anti-HSPA2 antibody. a Protein extract isolated from human testes was subjected to Western blot analysis. HSPA2 was detected with rabbit preimmune serum (left blot) or unpurified anti-HSPA2 rabbit polyclonal serum (right blot). b Protein extracts from human cell lines and human testes were subjected to Western blot analysis. We used proteins from HepG2 and A549 cell lines which previously were identified as not-transcribing and transcribing HSPA2 gene, respectively (Scieglinska et al. 2008). For HSPA2 detection antigen affinity purified anti-HSPA2 antibody was used. c Purified recombinant human HSPA1, HSPA2-GST, HSPA6 and HSPA8 proteins (0.5 μg each) were analyzed by Western blot. Left panel specific detection of HSPA2-GST protein with purified anti-HSPA2 antibody. Right panel commercially available anti-HSPA1 antibody (SPA-810) cross-reacts with HSPA6 protein. d Immunohistochemical detection of HSPA2 protein in testes. 1 example of testis tissue spot stained with anti-HSPA2 antibody; 2 higher magnification, note strong HSPA2 staining in spermatocytes (red asterisk) and spermaitds (green triangles), no staining in spermatogonia (white arrow); 3 specificity of HSPA2 detection in testes (anti-HSPA2 antibody and peptide competition, for details see “Material and methods” section); 4 testis stained with anti-HSPA1 antibody. Bar 100 μm