ZO1 Recombinant Rabbit Monoclonal Antibody [PSH07-09]
cat.: HA722797
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Monkey, Cynomolgus monkey
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: PSH07-09
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.
Molecular weight: Predicted band size: 195 kDa
Isotype: IgG
Immunogen: Recombinant protein within human ZO1 aa 1,401-1,748.
Positive control: A431 cell lysate, 293T cell lysate, HepG2 cell lysate, HeLa cell lysate, U-2 OS cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, COS-1 cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, MCF7, human kidney tissue, mouse kidney tissue, rat kidney tissue.
Subcellular location: Cell membrane, Cell junction, tight junction, gap junction, Cell projection, podosome.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
1:1,000
1:500-1:2,000
1:500-1:1,000
Uniprot #: SwissProt: Q07157 Human | P39447 Mouse
Entrez Gene: 292994 Rat
Alternative names: Tight junction protein 1 Tight junction protein ZO-1 Tight junction protein ZO1 TJP1 zo-1 Zo1 ZO1_HUMAN Zona occludens 1 Zona occludens 1 protein Zona occludens protein 1 Zonula occludens 1 protein Zonula occludens protein 1
Images
HA722797_1.jpg Fig1: Western blot analysis of ZO1 on different lysates with Rabbit anti-ZO1 antibody (HA722797) at 1/1,000 dilution.

Lane 1: A431 cell lysate (20 µg/Lane)
Lane 2: 293T cell lysate (20 µg/Lane)
Lane 3: HepG2 cell lysate (20 µg/Lane)
Lane 4: HeLa cell lysate (20 µg/Lane)
Lane 5: U-2 OS cell lysate (20 µg/Lane)
Lane 6: NIH/3T3 cell lysate (20 µg/Lane)
Lane 7: C2C12 cell lysate (20 µg/Lane)
Lane 8: COS-1 cell lysate (20 µg/Lane)
Lane 9: Mouse testis tissue lysate (40 µg/Lane)
Lane 10: Rat testis tissue lysate (40 µg/Lane)

Predicted band size: 195 kDa
Observed band size: 250 kDa

Exposure time: 3 minutes; ECL: K1802;
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 (HA722797) 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.
HA722797_2.jpg Fig2: Immunocytochemistry analysis of MCF7 cells labeling ZO1 with Rabbit anti-ZO1 antibody (HA722797) at 1/2,000 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-ZO1 antibody (HA722797) at 1/2,000 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.
HA722797_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-ZO1 antibody (HA722797) 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 (HA722797) 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.
HA722797_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-ZO1 antibody (HA722797) 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 (HA722797) 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.
HA722797_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-ZO1 antibody (HA722797) 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 (HA722797) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.