Cingulin Recombinant Rabbit Monoclonal Antibody [PSH06-19]
cat.: HA722516
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Cell, IP, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH06-19 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 137 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within Human Cingulin aa 750-1000. |
| Positive control: | Jurkat cell lysate, 293T cell lysate, Caco-2 cell lysate, MCF7 cell lysate, HUVEC cell lysate, MCF7, A431, human small intestine tissue, mouse small intestine tissue, rat small intestine tissue, Jurkat. |
| Subcellular location: | Cell junction. Tight junction. |
| Recommended Dilutions:
WB IHC-P IF-Cell IP FC |
1:1,000 1:200-1:1,000 1:100 1-2μg/sample 1:1,000 |
| Uniprot #: | SwissProt: Q9P2M7 Human | P59242 Mouse Entrez Gene: 310655 Rat |
| Alternative names: | 6330408J11Rik AI647528 AI987749 cgn CING_HUMAN Cingulin DKFZp779N1112 FLJ39281 KIAA1319 MGC118157 |
Images
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Fig1:
Western blot analysis of Cingulin on different lysates with Rabbit anti-Cingulin antibody (HA722516) at 1/1,000 dilution. Lane 1: Jurkat cell lysate Lane 2: 293T cell lysate Lane 3: Caco-2 cell lysate Lane 4: MCF7 cell lysate Lane 5: HUVEC cell lysate Lane 6: HeLa cell lysate (low expression) Lysates/proteins at 20 µg/Lane. Predicted band size: 137 kDa Observed band size: 137 kDa Exposure time: 30 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 (HA722516) at 1/1,000 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. |
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Fig2:
Immunocytochemistry analysis of MCF7 cells labeling Cingulin with Rabbit anti-Cingulin antibody (HA722516) at 1/100 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-Cingulin antibody (HA722516) at 1/100 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. |
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Fig3:
Immunocytochemistry analysis of A431 cells labeling Cingulin with Rabbit anti-Cingulin antibody (HA722516) at 1/100 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-Cingulin antibody (HA722516) at 1/100 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. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded human small intestine tissue with Rabbit anti-Cingulin antibody (HA722516) 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722516) 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. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-Cingulin antibody (HA722516) 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 (HA722516) 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. |
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Fig6:
Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Rabbit anti-Cingulin antibody (HA722516) 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722516) 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. |
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Fig7:
Flow cytometric analysis of Jurkat cells labeling Cingulin. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722516, 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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Fig8:
Cingulin was immunoprecipitated from 0.2 mg Jurkatcell lysate with HA722516 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722516 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Jurkat cell lysate (input) Lane 2: HA722516 IP in Jurkat cell lysate Lane 3: Rabbit IgG instead of HA722516 in Jurkat cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 32 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.