BLBP detected on rat embryo day 16 spinal cord T/S on 5 micron thick formalin-fixed paraffin wax embedded sections using Rabbit polyclonal to BLBP (ab32423; incubated at 1/200 for 16h at RT). 1% BSA (10 mins at RT) used as blocking agent. Secondary Antibody was biotin labeled anti-rabbit IgG (1/300). Immunoreactivity was visualised by standard streptAvidin-biotin-complex using peroxidase as a reporter and DAB as chromogen following citric acid (pH 6) heat mediated antigen retrieval. Expected specific staining observed with ab32423 with low signal to noise ratio.

Anti-BLBP antibody (ab32423) at 1/250 dilution + Mouse brain tissue lysate - total protein (0 days) (ab7188) at 10 µg

Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 15 kDa
Observed band size: 15 kDa

ICC/IF image of ab32423 stained rat PC12 cells. The cells were PFA fixed (10 min), permabilised in PBS-T (20 min) and incubated with the antibody (ab32423, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).

BLBP detected on mouse embryo day 16 spinal cord T/S on 5 micron thick formalin-fixed paraffin wax embedded sections using Rabbit polyclonal to BLBP (ab32423; incubated at 1/300 for 16h at RT). 1% BSA (10 mins at RT) used as blocking agent. Secondary Antibody was biotin labeled anti-rabbit IgG (1/300). Immunoreactivity was visualised by standard streptAvidin-biotin-complex using peroxidase as a reporter and DAB as chromogen following citric acid (pH 6) heat mediated antigen retrieval. Expected specific staining observed with ab32423.

Immunohistochemistical staining (Formaldehyde/PFA-fixed paraffin-embedded sections) for BLBP antibody (ab32423) on Zebrafish Tissue sections (young adult whole body T/S). Antigen retrieval step: Heat mediated Blocking step: 1% BSA for 10 mins at RT.

Primary Antibody: ab32423 incubated at 1/500 for 2 hours at RT.
Secondary Antibody: Biotin labelled goat anti rabbit IgG (1/300).

Immunohistochemistical detection (on Formaldehyde /PFA-fixed paraffin-embedded sections) of BLBP protein using BLBP antibody (ab32423) on Quail Tissue sections (developing spinal cord). Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT.

Primary Antibody: ab32423 incubated at 1/500 for 2 hours at RT.
Secondary Antibody: Biotin labelled goat anti rabbit IgG (1/300).