ET1610-3

S100 beta Recombinant Rabbit Monoclonal Antibody [SC57-02]

cat.: ET1610-3

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Zebrafish, Rat, Goat
Applications:	WB, IF-Cell, IF-Tissue, IP, IHC-P, IHC-Fr
Clonality:	Monoclonal
Clone number:	SC57-02

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 11 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within C-terminal human S100 beta.
Positive control:	Mouse brain tissue, mouse hippocampus tissue, rat brain tissue, mouse liver tissue lysates, SK-MEL-28 cell lysate, Rat brain tissue lysate, zebrafish tissue lysates, A375.
Subcellular location:	Cytoplasm, Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P IP IHC-Fr	1:1,000-1:2,000 1:50-1:200 1:50-1:200 1:50-1:1,000 Use at an assay dependent concentration. 1:100
Uniprot #:	SwissProt: P04271 Human \| P50114 Mouse \| P04631 Rat
Alternative names:	NEF Protein S100 B Protein S100-B S 100 calcium binding protein beta chain S 100 protein beta chain S-100 protein beta chain S-100 protein subunit beta S100 S100 calcium binding protein beta (neural) S100 calcium-binding protein B S100 protein beta chain S100B S100B_HUMAN S100beta

Images

	Fig1: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-S100 beta antibody (ET1610-3) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1610-3) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig2: Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue with Rabbit anti-S100 beta antibody (ET1610-3) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1610-3) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig3: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-S100 beta antibody (ET1610-3) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1610-3) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig4: Western blot analysis of S100 beta on mouse liver tissue lysates with Rabbit anti-S100 beta antibody (ET1610-3) at 1/1,000 dilution. Lysates/proteins at 20 µg/Lane. Predicted band size: 11 kDa Observed band size: 11 kDa Exposure time: 2 minutes; 15% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-3) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
	Fig5: Western blot analysis of S100 beta on different lysates with Rabbit anti-S100 beta antibody (ET1610-3) at 1/2,000 dilution. Lane 1: SK-MEL-28 cell lysate (15 µg/Lane) Lane 2: Rat brain tissue lysate (20 µg/Lane) Predicted band size: 11 kDa Observed band size: 11 kDa Exposure time: 3 minutes 10 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-3) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.
	Fig6: Western blot analysis of S100 beta on zebrafish tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-3, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature. Predicted band size: 11 kDa Observed band size: 14 kDa

	Fig7: Immunofluorescence analysis of paraffin-embedded mouse hippocampus tissue labeling S100 beta with Rabbit anti-S100 beta antibody (ET1610-3) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1610-3, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig8: Immunofluorescence analysis of frozen mouse brain tissue with Rabbit anti-S100 beta antibody (ET1610-3) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1610-3, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig9: Immunocytochemistry analysis of A375 cells labeling S100 beta with Rabbit anti-S100 beta antibody (ET1610-3) at 1/200 dilution. Cells were fixed in ice-cold 100% methanol for 5 minutes, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-S100 beta antibody (ET1610-3) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Fig10: Flow cytometric analysis of A375 cells labeling S100 beta.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1610-3, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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