Application
Immunohistochemistry free floating
Sample
Rat Tissue sections (Brain)
Specification
Brain
Other product details
Dilution
1/100
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 20°C
Secondary antibody
Name
Non-Abcam antibody was used: anti-rabbit (Jackson 711-165-152/89484)
Host species: Donkey
Clonality: Polyclonal
Conjugation: Cy3 ®
Host species: Donkey
Clonality: Polyclonal
Conjugation: Cy3 ®
Dilution
1/500
Additional data
Additional Notes
Methods
Rat brain tissue was fixed by transcardial perfusion with 4% paraformaldehyde, 0.1% glutaraldehyde, in 0.1 M phosphate buffer (pH7.4). The brain was postfixed with 4% paraformaldehyde in 0.1 M phosphate buffer (pH7.4) overnight at 4 degrees and then cut into 50 µm-thick parasagittal sections with a vibrating microtome.
Free-floating sections were incubated with 1/100, 1/300, 1/1,000, 1/3,000, or 1/10,000-diluted rabbit primary antibody against prodynorphin (preprodynorphin ab11137, lot#GR32272-1) in phosphate-buffered saline (PBS) containing 0.3% Triton X-100 and 1% normal donkey serum. After several washes with PBS, the sections were incubated with Cy3-conjugated donkey antibody to rabbit IgG (Jackson, 1/500) in the same incubation buffer as above for 3 hours at room temperature.
Results and Comments
This antibody provided clear punctate perikaryal immunoreactivity in a few neurons in the rat neocortex when used at high concentration (1/100 or 1/300, but not 1/3,000 or 1/10,000). This immunoreactivity was in good agreement with that shown with ab10280 (guinea pig) and published antibody (guinea pig Li et al., 1997, J Comp Neurol 386:229-44). Signal to noise ratio was fairly good, so this antibody may be used for immunohistochemistry on paraformaldehyde-fixed floating brain sections. Note that the titre of this antibody is relatively low, and thus a high concentration of the antibody is required.
The image is an example of indirect immunofluorescence labeling in the rat neocortex with 1/100-diluted ab11137 antibody.
Rat brain tissue was fixed by transcardial perfusion with 4% paraformaldehyde, 0.1% glutaraldehyde, in 0.1 M phosphate buffer (pH7.4). The brain was postfixed with 4% paraformaldehyde in 0.1 M phosphate buffer (pH7.4) overnight at 4 degrees and then cut into 50 µm-thick parasagittal sections with a vibrating microtome.
Free-floating sections were incubated with 1/100, 1/300, 1/1,000, 1/3,000, or 1/10,000-diluted rabbit primary antibody against prodynorphin (preprodynorphin ab11137, lot#GR32272-1) in phosphate-buffered saline (PBS) containing 0.3% Triton X-100 and 1% normal donkey serum. After several washes with PBS, the sections were incubated with Cy3-conjugated donkey antibody to rabbit IgG (Jackson, 1/500) in the same incubation buffer as above for 3 hours at room temperature.
Results and Comments
This antibody provided clear punctate perikaryal immunoreactivity in a few neurons in the rat neocortex when used at high concentration (1/100 or 1/300, but not 1/3,000 or 1/10,000). This immunoreactivity was in good agreement with that shown with ab10280 (guinea pig) and published antibody (guinea pig Li et al., 1997, J Comp Neurol 386:229-44). Signal to noise ratio was fairly good, so this antibody may be used for immunohistochemistry on paraformaldehyde-fixed floating brain sections. Note that the titre of this antibody is relatively low, and thus a high concentration of the antibody is required.
The image is an example of indirect immunofluorescence labeling in the rat neocortex with 1/100-diluted ab11137 antibody.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Dr. Kouichi Nakamura
Verified customer
投稿 Mar 24 2011