Cells: rat astrocytes
cultured for 4 week after isolation
and differentiated with cAMP and hydrocortisone treatment for 1 day
Fixation: 4% paraformaldehyde
Permeabilization: 0.2% triton X-100 in PBS for 10 min
Blocking: 2% bovine serum albumin (BSA) in PBS for 20 min
Primary antibody: 0.002 mg/mL in 2% BSA, 0.05% triton X-100 in PBS 1h RT
Secondary antibody: 1:1000 goat anti- guinea pig Ig Alexa Fluor 488 (Invitrogen; A11073) in 2% BSA, 0.05% triton X-100 in PBS 1h RT
Numerous astrocytes contain the cytoplasmic calcium-binding protein S100β which can be revealed with appropriate primary antibodies from several host species. Among them, guinea pig-anti-S100β (Synaptic Systems, Göttingen, cat. no. 287004) was shown to be useful for the specific detection of a larger portion of astroglia in the brain of tenrecs (Mack et al 2018 Brain Struct Funct – Fig 2). Additionally, this immunoreagent enables the indirect immunofluorescence labelling of astroglial S100β also in rodent brains but requires highly species-specific secondary antibodies.
On July 4, 2019 Andrea E Toth wrote:
Cells: rat astrocytes
cultured for 4 week after isolation
and differentiated with cAMP and hydrocortisone treatment for 1 day
Fixation: 4% paraformaldehyde
Permeabilization: 0.2% triton X-100 in PBS for 10 min
Blocking: 2% bovine serum albumin (BSA) in PBS for 20 min
Primary antibody: 0.002 mg/mL in 2% BSA, 0.05% triton X-100 in PBS 1h RT
Secondary antibody: 1:1000 goat anti- guinea pig Ig Alexa Fluor 488 (Invitrogen; A11073) in 2% BSA, 0.05% triton X-100 in PBS 1h RT
On January 27, 2019 Wolfgang Härtig, University of Leipzig wrote:
Numerous astrocytes contain the cytoplasmic calcium-binding protein S100β which can be revealed with appropriate primary antibodies from several host species. Among them, guinea pig-anti-S100β (Synaptic Systems, Göttingen, cat. no. 287004) was shown to be useful for the specific detection of a larger portion of astroglia in the brain of tenrecs (Mack et al 2018 Brain Struct Funct – Fig 2). Additionally, this immunoreagent enables the indirect immunofluorescence labelling of astroglial S100β also in rodent brains but requires highly species-specific secondary antibodies.
I can highly recommend this antibody
Mack et al. 2018 Brain Struct Funct: https://link.springer.com/article/10.1007%2Fs00429-018-1730-1