Anti-S100 beta antibody [EP1576Y] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

This data was developed using the same antibody clone in a different buffer formulation (ab52642).

Immunohistochemical analysis of formalin fixed paraffin embedded human cerebrum labelling S100 beta with ab52642 at a concentration of 0.01 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). ab52642 anti-S100 beta antibody [EP1576Y] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

This data was developed using ab52642, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain tissue* labelling S100B with ab52642 at 1ug/ml followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining in human Alzheimer's brain.

The section was incubated with ab52642 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0 epitope retrieval solution1) for 20 mins.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

Immunofluorescent imaging of human native and acellular 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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

Image from Gerli MFM et al. PLoS One. 2018;13(1):e0191497 Fig 3.; doi: 10.1371/journal.pone.0191497.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

Clone EP1576Y (ab215989) has been successfully conjugated by Abcam. This image was generated using Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (Alexa Fluor^® 647). Please refer to ab196175 for protocol details.

ab196175 staining S100 beta in A375 cells. 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 ab196175 at 1/50 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

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

This product also gave a positive signal under the same testing conditions in A375 cells fixed with 4% formaldehyde (10 min).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

• IHC-P

AbReview37991****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

This image is courtesy of an Abreview submitted by Heiko Locher.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

This data was developed using the same antibody clone in a different buffer formulation (ab52642).

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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

Immunohistochemistry (Frozen sections) analysis of mouse cerebrum tissue sections labeling S100 beta with purified ab52642 at 1/100 (9.9 μg/ml) dilution. 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

Immunohistochemistry (Frozen sections) analysis of rat cerebrum tissue sections labeling S100 beta with purified ab52642 at 1/100 (9.9 μg/ml) dilution. 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

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 Alexa Fluor^® 488 (ab150077) at a dilution of 1/400.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

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.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

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

• WB

Lab

Western blot - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

This data was developed using ab52642, the same antibody clone in a different buffer formulation.

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

We recommend using a 14% SDS-PAGE gel to obtain a clear and strong S100β band.

Lanes 1 and 3:

Western blot - Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/s100-beta-antibody-ep1576y-astrocyte-marker-ab52642'>ab52642</a>) at 1/1000 dilution

Lanes 2 and 4:

Western blot - Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/s100-beta-antibody-ep1576y-astrocyte-marker-ab52642'>ab52642</a>) at 1/10000 dilution

All lanes:

A375 ( human malignant melanoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 10 kDa

false

Exposure time: 140s

• WB

Supplier Data

Western blot - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

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

All lanes:

Western blot - Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/s100-beta-antibody-ep1576y-astrocyte-marker-ab52642'>ab52642</a>) 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

false

• WB

Supplier Data

Western blot - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

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

All lanes:

Western blot - Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/s100-beta-antibody-ep1576y-astrocyte-marker-ab52642'>ab52642</a>) at 1/5000 dilution

All lanes:

B16F0 (mouse 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

false

• WB

Supplier Data

Western blot - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52642).

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

All lanes:

Western blot - Anti-S100 beta antibody [EP1576Y] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/s100-beta-antibody-ep1576y-astrocyte-marker-ab52642'>ab52642</a>) at 1/10000 dilution

Lane 1:

Mouse spinal cord at 20 µg

Lane 2:

Rat brain at 20 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

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-S100 beta antibody [EP1576Y] - BSA and Azide free (AB215989)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.