Anti-S100 Beta Antibody - Astrocyte Marker (ab52642)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (ab52642)

S100 beta is present in CNV lesions.

A) S100 beta expression in a normal WT mouse retina. Strong immunoreactivity is present in the astrocytes (arrow). The position of the inner nuclear layer (INL) and outer nuclear layer (ONL) are indicted. Scale bar is 50 μm.

B) S100 beta expression in WT mouse retinal at day 7 post-laser treatment. S100B was detected in the outer plexiform layer (arrowheads). Strong S100 beta expression was detected at the site of CNV. Scale bar is 50 μm.

Image from Chen M et al. PLoS One. 2014;9(2):e89548. Fig 3.; doi: 10.1371/journal.pone.0089548.

S100 beta expression in normal and CNV-affected mouse retinas, highlighting astrocyte reactivity and increased expression in the outer plexiform layer.

Immunocytochemistry/ Immunofluorescence - Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (ab52642)

ab52642 staining S100 beta in primary rat neurons/glia, DIV14 (prepared from E18 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDHEP). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab52642 at 5μg/ml and ab195883, Rat monoclonal to alpha Tubulin (Alexa Fluor® 488), at 2μg/ml (shown in green). The secondary antibody (shown in red) was ab150083, Alexa Fluor® 647 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h at room temperature. Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Confocal microscopy image of primary rat neurons/glia stained for S100 beta (red), alpha Tubulin (green), and DNA (blue).

Western blot - Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (ab52642)

Blocking and Diluting buffer and concentration: 5% NFDM/TBST

All lanes:

Western blot - Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (ab52642) at 1/5000 dilution

All lanes:

A-375 (human malignant melanoma cell line) at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 10 kDa

Observed band size: 11 kDa

Western blot of A-375 cells stained with anti-S100 beta antibody, showing a band at approximately 11 kDa.

Immunocytochemistry/ Immunofluorescence - Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (ab52642)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 A-375 (human malignant melanoma cell line) cells labeling S100 beta with purified ab52642 at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 488) ab150077 secondary antibody at 1/500 dilution (green). The nuclear counter stain is DAPI (blue). The negative control is as follows;

ab52642 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.

Immunofluorescence image of human melanoma cells stained for S100 beta protein (green) and nuclei (blue). Negative control is also shown.

Immunohistochemistry (Frozen sections) - Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (ab52642)

IHC image of S100 beta staining in a section of frozen normal human cerebral cortex performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab16659, 1/5000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

IHC image of S100 beta staining in normal human cerebral cortex.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (ab52642)

Immunohistochemical analysis of embryonic mouse brain tissue, staining S100 beta with unpurified ab52642.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Image from Selvadurai HJ & Mason JO. PLoS One. 2011;6(8):e23012. Epub 2011 Aug 8. Fig 5.; doi:10.1371/journal.pone.0023012; August 8 2011 PLoS ONE 6(8): e23012.

Image of mouse brain tissue stained with S100 beta antibody to visualize specific proteins.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (ab52642)

Immunohistochemical analysis of paraffin embedded human cerebral cortex tissue labeling S100 beta with purified ab52642 at 1/1000 dilution. Secondary antibody was Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Counter stain: Hematoxylin.

Negative control: Using PBS instead of primary antibody.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

S100 beta protein expression in human cerebral cortex tissue