ZO-1/TJP1 Rabbit Polyclonal Antibody

• Catalog Number: orb499587
• Category: Antibodies
• Description: ZO-1/TJP1 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: WB
• Predicted Reactivity: Bovine, Canine, Gallus, Goat, Guinea pig, Mouse, Porcine, Rat, Sheep
• Reactivity: Human
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human ZO-1 (1551-1702/1733aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 191 kDa
• Target: TJP1
• UniProt ID: Q07157
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: ZO1_HUMAN; Tight junction protein ZO-1; TJP1; ZO1;
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Black line: Positive blank control (293T), Negative blank control (HL60), Green line: Primary Antibody (Rabbit Anti-ZO-1 antibody), Orange line: Isotype Control Antibody (Rabbit IgG). Blue line: Secondary Antibody (Goat anti-rabbit IgG-AF647), 293T (Positive) and HL60 (Negative control) cells (black) were fixed with 4% PFA for 10 min at room temperature, permeabilized with PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with ZO-1 Antibody (orb499587) at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody (blue) incubation for 40 min at room temperature. Acquisitions of 20000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

Blank control: 293T Cells (blue). Primary Antibody: Rabbit Anti-ZO-1/AF647 Conjugated antibody, dilution: 1 µg in 100 µl 1X PBS containing 0.5% BSA, Isotype Control Antibody: Rabbit IgG/AF647 (orange), used under the same conditions. Protocol, The cells were washed twice with phosphate-buffered saline (PBS). The cells were incubated in 1X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions followed by the antibody (1 µg/1x10^6 cells) for 30 min on ice. Acquisition of 20000 events was performed.

Independently Validated Antibody, Formalin-fixed and paraffin embedded human testis labeled with Anti-ZO-1 Polyclonal Antibody, Unconjugated (orb499587) at 1:250 followed by conjugation to the secondary antibody.

Sample: Lane 1: Huvec (Human) Cell Lysate at 30 ug, Lane 2: A431 (Human) Cell Lysate at 30 ug, Primary: Anti-ZO-1 (orb499587) at 1/1000 dilution, Anti-beta-Actin (orb500817) at 1/2000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 220 kD, Observed band size: 220 kD.