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Western blot analysis of Zona Occludens protein 1 (ZO-1) was performed by loading 50ug of MDCK (lane 2) and NRK (lane 3) cell lysates and 2ul SeeBlue® Plus2 Prestained Protein Ladder (Product # LC5925) (lane 1) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 1% BSA/TBST for at least 1 hour at room temperature. ZO-1 was detected using a monoclonal antibody (Product # 33-9100) at a concentration of 1ug/ml in blocking buffer overnight at 4°C on a rocking platform, followed by a goat anti-mouse IgG Alexa Fluor 790 conjugated secondary antibody (Product # A11357) at a dilution of 1:10,000 for at least 1 hour. Fluorescent detection was performed using the Odyssey® CLx imaging system (Li-cor Biosciences). Images generated by Joell Solan in Paul Lampe Lab at Fred Hutchinson Cancer Research Center.
|Tested species reactivity||Mouse|
|Host / Isotype||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||Alexa Fluor® 790|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against bovine IgG, goat IgG, rabbit IgG, rat IgG, human IgG and human serum|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1-10 µg/mL|
|Immunocytochemistry (ICC)||1-10 µg/mL|
|Immunofluorescence (IF)||1-10 µg/mL|
|Western Blot (WB)||1:10,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This secondary antibody is designed for fluorescent Western blot detection on various near-infrared fluorescence instruments. This antibody can be used for multi-color and multiplexing detection when using other antibodies conjugated to compatible Alexa Fluor™ dyes and wavelengths. Other applications of this antibody include immunofluorescent and fluorescent imaging applications when using instrumentation with appropriate excitation and detection capabilities.
Anti-Mouse secondary antibodies are affinity-purified antibodies with well-characterized specificity for mouse immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.