Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain)
Specification
Brain
Other product details
Dilution
1/100
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 20°C · Diluent: PBST
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-rabbit Alexa Fluor 488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Dilution
1/1000
Additional data
Additional Notes
This antibody produced a cytoplasmic neuronal staining in the suprachiasmatic nucleu and in some glial cells stained in many brain areas. In the hippocampus, the staining is located in thin terminals.
Pre-absorption experiment with a 10X exces of peptide ab 17041, which is a peptide of the peptide 123-130 or peptide ab17030, which is the phosphorylation site Y125. The pictures on the below column show the staining of the pre-absorption experiment without peptide (left) and with the two different peptides (ab17041 peptide 125-130) or ab17030 (peptide Y125). It shows that with the 125-130 peptide, the staining in the terminals has disappeared, while with the Y125, all the stainings have disappeared. This suggests that the staining obtained is specific for this phosphorylation site of the alpha Synuclein. Pictures on the below colums were taken at the same level of the hippocampus. The arrows show the stainings in glial or neuroglial form cells, while the stars locate the area where the punctiform staining is, or should be located.
The sections used came from animals perfused fixed with Paraformaldehyde 4% with 15% of a solution of saturated picric acid, in phosphate buffer 0.1M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Brains were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.
Pre-absorption experiment with a 10X exces of peptide ab 17041, which is a peptide of the peptide 123-130 or peptide ab17030, which is the phosphorylation site Y125. The pictures on the below column show the staining of the pre-absorption experiment without peptide (left) and with the two different peptides (ab17041 peptide 125-130) or ab17030 (peptide Y125). It shows that with the 125-130 peptide, the staining in the terminals has disappeared, while with the Y125, all the stainings have disappeared. This suggests that the staining obtained is specific for this phosphorylation site of the alpha Synuclein. Pictures on the below colums were taken at the same level of the hippocampus. The arrows show the stainings in glial or neuroglial form cells, while the stars locate the area where the punctiform staining is, or should be located.
The sections used came from animals perfused fixed with Paraformaldehyde 4% with 15% of a solution of saturated picric acid, in phosphate buffer 0.1M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Brains were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.
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. Sophie Pezet
Verified customer
投稿 Nov 03 2011