ICC/IF image of ab32481 stained mouse embryonic stem cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32481, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).
Western blot - BDP / ARID3B antibody (ab32481)
All lanes : Anti-BDP / ARID3B antibody (ab32481) at 1 µg/ml
Lane 1 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate Lane 2 : Thyroid Gland (Mouse) Tissue Lysate Lane 3 : Testis (Mouse) Tissue Lysate Lane 4 : Thymus (Mouse) Tissue Lysate Lane 5 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate Lane 6 : Cerebellum (Mouse) Tissue Lysate Lane 7 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
IHC image of ab32481 staining in mouse testes formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32481, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.