Anti-ZO1 tight junction protein antibody [mAbcam 61357] (ab61357)
Key features and details
- Mouse monoclonal [mAbcam 61357] to ZO1 tight junction protein
- Suitable for: Flow Cyt, WB, IP
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-ZO1 tight junction protein antibody [mAbcam 61357]
See all ZO1 tight junction protein primary antibodies -
Description
Mouse monoclonal [mAbcam 61357] to ZO1 tight junction protein -
Host species
Mouse -
Tested applications
Suitable for: Flow Cyt, WB, IPmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Dog -
Immunogen
Synthetic peptide corresponding to Human ZO1 tight junction protein aa 750-850 conjugated to keyhole limpet haemocyanin.
(Peptide available asab93087) -
Positive control
- This antibody gave a positive signal in the following whole cell lysates: HepG2; HEK293; A431; Caco2.
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General notes
This antibody clone is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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Purity
IgG fraction -
Clonality
Monoclonal -
Clone number
mAbcam 61357 -
Myeloma
Sp2/0-Ag14 -
Isotype
IgG1 -
Research areas
Images
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All lanes : Anti-ZO1 tight junction protein antibody [mAbcam 61357] (ab61357) at 5 µg/ml
Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 4 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 195 kDa
Observed band size: 240 kDa why is the actual band size different from the predicted?
Additional bands at: 38 kDa, 44 kDa, 48 kDa, 85 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 30 seconds
The 240-kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to ZO1 tight junction protein. -
ZO1 tight junction protein was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Mouse monoclonal to and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab61357.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.
Band: 240kDa, non specific band - 90kDa: We are unsure as to the identity of this extra band; ZO1 tight junction protein
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Overlay histogram showing MCF-7 cells stained with ab61357 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab61357, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.