SCGN Recombinant Rabbit Monoclonal Antibody [JE60-48]
cat.: HA721891
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, FC
Clonality: Monoclonal
Clone number: JE60-48
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. 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: 32 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human SCGN aa 227-276 / 276.
Positive control: TT cell lysate, human brain tissue lysate, mouse eyeball tissue lysate, mouse pancreas tissue lysate, rat eyeball tissue lysate, TT, human brain tissue, human colon tissue, human stomach cancer tissue, mouse retina tissue, rat retina tissue.
Subcellular location: Cytoplasm, Cytoplasmic vesicle, Membrane, Secreted.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC

1:2,000
1:500-1:2,000
1:200
1:1,000
Uniprot #: SwissProt: O76038 Human | Q91WD9 Mouse | Q6R556 Rat
Alternative names: Calbindin like CALBL DJ501N12.8 OTTHUMP00000016124 Scgn SECRET Secretagogin Secretagogin EF hand calcium binding protein SEGN SEGN_HUMAN Setagin
Images
HA721891_1.jpg Fig1: Western blot analysis of SCGN on different lysates with Rabbit anti-SCGN antibody (HA721891) at 1/2,000 dilution.

Lane 1: TT cell lysate
Lane 2: Human brain tissue lysate
Lane 3: Mouse eyeball tissue lysate
Lane 4: Mouse pancreas tissue lysate
Lane 5: Rat eyeball tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 32 kDa
Observed band size: 32 kDa

Exposure time: 5 minutes;

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 (HA721891) 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.
HA721891_2.jpg Fig2: Immunocytochemistry analysis of TT cells labeling SCGN with Rabbit anti-SCGN antibody (HA721891) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SCGN antibody (HA721891) 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.
HA721891_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-SCGN antibody (HA721891) at 1/2,000 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 (HA721891) at 1/2,000 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.
HA721891_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-SCGN antibody (HA721891) at 1/2,000 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 (HA721891) at 1/2,000 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.
HA721891_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue with Rabbit anti-SCGN antibody (HA721891) at 1/2,000 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 (HA721891) at 1/2,000 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.
HA721891_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse retina tissue with Rabbit anti-SCGN antibody (HA721891) at 1/2,000 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 (HA721891) at 1/2,000 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.
HA721891_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat retina tissue with Rabbit anti-SCGN antibody (HA721891) at 1/2,000 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 (HA721891) at 1/2,000 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.
HA721891_8.jpg Fig8: Flow cytometric analysis of TT cells labeling SCGN.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721891, 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.