polyclonal guinea pig anti-calbindin  (Synaptic Systems)

Journal: eLife

Article Title: Endosomal dysfunction contributes to cerebellar deficits in spinocerebellar ataxia type 6

doi: 10.7554/eLife.90510

Figure Lengend Snippet: List of antibodies.

Article Snippet: Calbindin , Anti-calbindin polyclonal (guinea pig) , Synaptic Systems , 214 004 , 1:500 , 2, 3, 6, 9.

Techniques:

Journal: eLife

Article Title: Synergism of type 1 metabotropic and ionotropic glutamate receptors in cerebellar molecular layer interneurons in vivo

doi: 10.7554/eLife.56839

Figure Lengend Snippet: ( A ) Two-photon laser scanning microscopy (2PLSM) image of the outermost cerebellar molecular layer in vivo shows cytoplasmic GCaMP3 and parasagittally aligned MLI neurites in a horizontal plane of focus. ( B ) Confocal stack projections of sagittal cerebellar slices from mice expressing AAV2/1hSyn.Flex.GCaMP3 on the background of PV-promoter driven CRE recombinase. Left : slice stained with an antibody against calbindin, a protein expressed in PCs and not in MLIs. Note that calbindin positive PCs do not express GCaMP3. Right : slice stained with an antibody against PV, a protein expressed in both PCs and MLIs. Note the colocalization between GCaMP3 and PV positive MLIs. ( C ) A beam pattern of Ca i rise is illustrated by the ΔF/Fo image at the peak of the fluorescence increase evoked in the molecular layer by a 0.5 s long PF stimulation at 100 Hz (horizontal plane of focus) using a bi-polar theta glass pipette as depicted in the cartoon. The highest value for the pseudo-color scale is 100%. ( D ) Time course of the somatic MLI Ca i signals evoked in a representative MLI by 100 Hz PF stimulations of various durations. The arrow indicates the time of stimulation onset. The inset shows an average of pre-stimulus images to illustrate the ROI analyzed. ( E ) Average peak ΔF/Fo values from 18 somata, normalized to the peak value for 200 ms trains at 100 Hz.

Article Snippet: The slices were first incubated in PBS containing 0.3% Triton and 10% fetal bovine serum for 5 hr at room temperature then at 4°C overnight with a mixture of rabbit polyclonal serum anti-Parvalbumin (PV25, SWANT, Marly, Switzerland) and guinea pig polyclonal serum anti calbindin D28k (Synaptic Systems, Gôttingen, Germany) diluted to 1/1000 in PBS containing 1 mg/ml of Bovine Serum Albumine (BSA).

Techniques: Laser-Scanning Microscopy, In Vivo, Expressing, Staining, Fluorescence, Transferring