5-HT (Serotonin) Recombinant Rabbit Monoclonal Antibody [PSH06-04]
cat.: HA722503
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Species independent
Applications: WB, IHC-P, IF-Tissue, IHC-Fr, ELISA
Clonality: Monoclonal
Clone number: PSH06-04
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Isotype: IgG
Immunogen: 5-HT-OVA
Recommended Dilutions:
  ELISA
  WB
  IHC-P
  IF-Tissue
  IHC-Fr
1:1,000
1:2,000
1:200-1:1,000
1:200
1:200
Alternative names: 5 hydroxytryptamine 5HT
Images
HA722503_1.jpg Fig1: Western blot analysis of 5-HT (Serotonin) on different peptides with Rabbit anti-5-HT (Serotonin) antibody (HA722503) at 1/2,000 dilution.

Lane 1: 5-HT-BSA (positive)
Lane 2: Tryptamine-BSA (negative)
Lane 3: 5-Methoxytryptamine-BSA (negative)
Lane 4: Dopamine-BSA (negative)
Lane 5: BSA (negative)

Lysates/proteins at 30 ng/Lane.

Predicted band size: 66 kDa
Observed band size: 66 kDa

Exposure time: 1 minute; 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 (HA722503) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA722503_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-5-HT (Serotonin) antibody (HA722503) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA722503) at 1/200 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.
HA722503_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human small intestine tissue with Rabbit anti-5-HT (Serotonin) antibody (HA722503) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA722503) 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.
HA722503_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-5-HT (Serotonin) antibody (HA722503) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA722503) 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.
HA722503_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Rabbit anti-5-HT (Serotonin) antibody (HA722503) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA722503) 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.
HA722503_6.jpg Fig6: Immunofluorescence analysis of paraffin-embedded human small intestine tissue labeling 5-HT (Serotonin) with Rabbit anti-5-HT (Serotonin) antibody (HA722503) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA722503, 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).
HA722503_7.jpg Fig7: Immunofluorescence analysis of paraffin-embedded mouse small intestine tissue labeling 5-HT (Serotonin) with Rabbit anti-5-HT (Serotonin) antibody (HA722503) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA722503, 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).
HA722503_8.jpg Fig8: Immunofluorescence analysis of paraffin-embedded rat small intestine tissue labeling 5-HT (Serotonin) with Rabbit anti-5-HT (Serotonin) antibody (HA722503) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA722503, 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).
HA722503_9.jpg Fig9: Immunofluorescence analysis of frozen mouse small intestine tissue with Rabbit anti-5-HT (Serotonin) antibody (HA722503) at 1/200 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 (HA722503, 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).
HA722503_10.jpg Fig10: Immunofluorescence analysis of frozen rat small intestine tissue with Rabbit anti-5-HT (Serotonin) antibody (HA722503) at 1/200 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 (HA722503, 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).
HA722503_11.jpg Fig11: Competitive ELISA analysis of 5-HT was performed by coating wells of a 96-well plate with 50 µL per well of 5-HT-BSA diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 1%BSA blocking buffer, and incubated with 50 µL per well of 5-HT monoclonal antibody at concentration of 1 µg/mL with serial diluted 5-HT starting from a concentration of 10µg/mL for 45 min at 37℃. The plate was washed and incubated with 50 µL per well of an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:70,000 for 30 min at 37℃. Detection was performed using an Ultra TMB Substrate for 10 minutes at 37℃ in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.