Recombinant 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). 

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

IHC image of S100 beta staining in a section of frozen normal human cerebral cortex performed on a Leica BOND^TM 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.

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.

S100 beta was immunoprecipitated from human fetal brain with purified ab52642 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab52642 and Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as secondary antibody at 1/1000 dilution.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

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.

Immunohistochemistry (Frozen sections) analysis of rat cerebrum tissue sections labeling S100 beta with purified ab52642 at 1/100 (9.9 µg/ml). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. DAPI was used as a counterstain.

Immunohistochemistry (Frozen sections) analysis of mouse cerebrum tissue sections labeling S100 beta with purified ab52642 at 1/100 (9.9 µg/ml). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. DAPI was used as a counterstain.

Immunohistochemical analysis of paraffin embedded rat 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 antibody ab52642 was used with Tissue Clearing Kit ab243298 to penetrate, stain and clear a 1 mm coronal section of mouse brain. Blue: DAPI, Green: S100 beta.

Learn more about tissue clearing kits, reagents, and protocols designed to make it easier to stain thick tissue sections and get more data from each valuable tissue section.

To use this antibody with tissue clearing, use Tissue Clearing Kit ab243298. For 1 mm brain sections, we recommend a starting dilution of 1:200, and also using Goat Anti-Rabbit IgG H&L AlexaFluor488 (ab150077) at a dilution of 1:400.

Immunohistochemical analysis of paraffin embedded mouse 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 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.

Immunofluorescent imaging of native and acellular human peripheral nerve sections stained for the axon protein Neurofilaments (NF200), the Schwann’s cell marker S100β (ab52642) and for the extracellular matrix proteins Laminin and Collagen IV. Sections were counterstained with DAPI to confirm the removal of the cell nuclei upon decellularization (scale bar: 100 μm).

Unpurified ab52642 staining S100 beta in human spiral ganglion tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 1% BSA for 30 minutes at room temperature; antigen retrieval was by heat mediation in citrate buffer, pH 6.0. Samples were incubated with primary antibody (1/200 in PBS-T + 1% BSA) for 12 hours. An Alexa Fluor^® 488-conjugated Donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

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

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

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.