ICC/IF image of ab6319 stained SKNSH cells. The cells were 4% formaldehyde 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 ab6319 at 10µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- mouse (ab150117) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

All lanes : Anti-CNPase antibody [11-5B] (ab6319) at 1/100 dilutionLane 1 : Spinal Cord (Human) Tissue Lysate - adult normal tissue (ab29188)Lane 2 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)Lane 3 : Spinal Cord (Mouse) Tissue LysateLane 4 : Brain (Mouse) Tissue Lysate Lane 5 : Spinal Cord (Rat) Tissue LysateLane 6 : Brain (Rat) Tissue LysateLysates/proteins at 20 µg per lane.SecondaryGoat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

IHC-P image of CNPase staining on rat brain sections using ab6319 (1:1600). heat mediated antigen retrieval on paraffin embedded sections was performed using citric acid. The sections were then blocked with 1% BSA for 10 min at 21°C. The primary antibody was incubated for 16 hours at 21°C. The sections were then incubated in Goat anti-mouse (Biotin) at 1:200. See Abreview

ab6319 at a 1/2000 dilution staining mouse brain tissue sections by Immunohistochemistry (Frozen sections). The tissue was paraformaldehyde fixed and blocked with 2% BSA. The antibody was incubated with the tissue for 10 hours and then bound antibody was detected using a biotinylated goat anti-mouse antibody.CNPase staining in the striatum is shown in the image.This image is courtesy of an Abreview submitted by Bing Lang on 19 April 2006.See Abreview

ab6319 at a 1/200 dilution staining rat spinal cord cells by ICC/IF. The cells were ethanol fixed and blocked with 5% serum. The antibody was incubated with the cells for 1 hour and then bound antibody was detected using an Alexa Fluor® 488 conjugated Goat anti-mouse antibody. This image is courtesy of an Abreview submitted by Nancy Nutile-McMenemy.See Abreview

ab6319 at a 1/200 dilution staining rat spinal cord tissue sections from a 4% PFA transcardially perfused animal by Immunohistochemistry (Frozen sections). The tissue was paraformaldehyde fixed and incubated with the antibody for 18 hours. Bound antibody was detected using an HRP conjugated goat anti-mouse polyclonal antibody. This image is courtesy of an Abreview submitted by Nancy Nutile-McMenemy.See Abreview

All lanes : Anti-CNPase antibody [11-5B] (ab6319) at 1/500 dilutionLane 1 : 40ug rat brain homegenate (whole tissue lysate) animal #661Lane 2 : 40ug rat brain homegenate (whole tissue lysate) animal #655Lane 3 : 40ug rat brain homegenate (whole tissue lysate) animal #668Lane 4 : 40ug rat brain homegenate (whole tissue lysate) animal #662Lane 5 : 40ug rat brain homegenate (whole tissue lysate) animal #659Lane 6 : 40ug rat brain homegenate (whole tissue lysate) animal #670SecondaryHRP conjugated goat anti-mouse antibodydeveloped using the ECL techniquePerformed under reducing conditions.

All lanes : Anti-CNPase antibody [11-5B] (ab6319) at 1/750 dilutionLane 1 : Spinal Cord homogenate (whole tissue lysate)Lane 2 : Spinal Cord homogenate (whole tissue lysate)Lane 3 : Spinal Cord homogenate (whole tissue lysate)Lysates/proteins at 2 µg per lane.SecondaryHRP conjugated sheep anti-mouse IgG

ab6319 staining mouse brain tissue sections by IHC-FoFr.  Sections were PFA fixed and permeabilized in 0.1% Triton X-100 prior to blocking in 0.5% TNB for 30 minutes at 25°C. The primary antibody was diluted 1/250 and incubated with the sample for 18 hours at 25°C.  An Alexa Fluor® 488 conjugated goat anti-mouse antibody, diluted 1/250, was used as the secondary.Image demonstrates a 2-D depth projection through the superficial cortex.See Abreview

ab6319 staining CNPase in rat oligodendrocytes by Immunocytochemistry/ Immunofluorescence.Cells were fixed in paraformaldehyde, blocked using 5% serum for 10 minutes at 25°C, then incubated with ab6319 at a 1/200 dilution for 2 hours at 25°C. The secondary used was a goat anti-mouse Cy3 conjugated polyclonal at a 1/100 dilution.See Abreview

Overlay histogram showing SH-SY5Y cells stained with ab6319 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab6319, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.