Western blot - Anti-CNPase antibody [11-5B] - Oligodendrocyte Marker (ab6319)

All lanes : Anti-CNPase antibody [11-5B] - Oligodendrocyte Marker (ab6319) at 1/100 dilution

Lane 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 Lysate
Lane 4 : Spinal Cord (Mouse) Tissue Lysate
Lane 5 : Spinal Cord (Rat) Tissue Lysate
Lane 6 : Brain (Rat) Tissue Lysate

Lysates/proteins at 20 µg per lane.

Secondary
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 48 kDa


Exposure time : 1 minute

This antibody was raised against full length native CNPase and is predicted to recognize both isoforms. The predicted molecular weights of isoforms CNPI and CNPII are 45- and 48-kDa respectively.

Immunohistochemistry (Frozen sections) - CNPase antibody [11-5B] - Oligodendrocyte Marker (ab6319)

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.

Immunocytochemistry/ Immunofluorescence - CNPase antibody [11-5B] - Oligodendrocyte Marker (ab6319)

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.

Immunohistochemistry (Frozen sections) - CNPase antibody [11-5B] - Oligodendrocyte Marker (ab6319)

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.

Western blot - CNPase antibody [11-5B] - Oligodendrocyte Marker (ab6319)

All lanes : Anti-CNPase antibody [11-5B] - Oligodendrocyte Marker (ab6319) at 1/500 dilution

Lane 1 : 40ug rat brain homegenate (whole tissue lysate) animal #661
Lane 2 : 40ug rat brain homegenate (whole tissue lysate) animal #655
Lane 3 : 40ug rat brain homegenate (whole tissue lysate) animal #668
Lane 4 : 40ug rat brain homegenate (whole tissue lysate) animal #662
Lane 5 : 40ug rat brain homegenate (whole tissue lysate) animal #659
Lane 6 : 40ug rat brain homegenate (whole tissue lysate) animal #670

Secondary
HRP conjugated goat anti-mouse antibody
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 48 kDa
Observed band size : 48 kDa


Exposure time : 5 minutes

This image is courtesy of an Abreview submitted by Ms Nancy Nutile-McMenemy

Western blot - CNPase antibody [11-5B] - Oligodendrocyte Marker (ab6319)

All lanes : Anti-CNPase antibody [11-5B] - Oligodendrocyte Marker (ab6319) at 1/750 dilution

Lane 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.

Secondary
HRP conjugated sheep anti-mouse IgG

Predicted band size : 48 kDa
Observed band size : 45,47 kDa (why is the actual band size different from the predicted?)

This image is courtesy of an anonymous Abreview

Immunohistochemistry (PFA perfusion fixed frozen sections) - CNPase antibody [11-5B] - Oligodendrocyte Marker (ab6319)

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.

This image is courtesy of an anonymous Abreview

Immunocytochemistry/ Immunofluorescence - CNPase antibody [11-5B] - Oligodendrocyte Marker (ab6319)

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.

Image courtesy of an anonymous Abreview.