61-7300

antibody from Invitrogen Antibodies

Targeting: TJP1 DKFZp686M05161, MGC133289, ZO-1

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ELISA

Immunoprecipitation

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Validation data

Product number: 61-7300 - Provider product page
Provider: Invitrogen Antibodies
Product name: ZO-1 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Other
Description: 61-7300 has been successfully used in Western blot, Immunoprecipitation, Immunofluorescence, ELISA and Immunohistochemistry. To achieve appropriate staining results on formalin-fixed, paraffin embedded tissue sections, protease Type XIV (Actinase E) pretreatment is required. The recommended dilution and incubation for protease is 1-2 mg/mL at 37°C for 10 minutes. Other enzyme and Heat Induced Epitope Retrieval pre-treatment methods may not be suitable for this particular antibody. Frozen sections have been successfully stained after fixation with acetone at 4°C.
Reactivity: Human, Mouse, Rat, Canine, Guinea Pig
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 0.25 mg/mL
Storage: -20°C

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of ZO-1 (green) in Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes and blocked with 3% Blocker BSA (Product # 37525) in PBS for 15 minutes at room temperature. Cells were stained with or without ZO-1 rabbit polyclonal antibody (Product # 61-7300), at a concentration of 5 µg/mL for 1 hour at room temperature, and then incubated with a Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:1000 for 1 hour at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of ZO-1 (green) in Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes and blocked with 3% Blocker BSA (Product # 37525) in PBS for 15 minutes at room temperature. Cells were stained with or without ZO-1 rabbit polyclonal antibody (Product # 61-7300), at a concentration of 5 µg/mL for 1 hour at room temperature, and then incubated with a Goat anti-Rabbit IgG (Heavy Chain) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:1000 for 1 hour at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of ZO-1 was performed using 70% confluent log phase Caco-2 and Jurkat cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with ZO-1 Polyclonal Antibody (Product # 61-7300) at 1 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 488 (Product # A32790, 1:2,000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image of Caco-2, which is a high expressing model for ZO-1 showing junctional localization. Panel e represents the merged image of Jurkat cells that are a low expressing model for ZO-1. Panel f represents control cells with no primary antibody to assess background. The images were captured at 40x magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of ZO-1 was performed using 70% confluent log phase MDCK cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with ZO-1 Polyclonal Antibody (Product # 61-7300) at 1 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 488 (Product # A32790, 1:2,000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image of MDCK showing junctional localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40x magnification.