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Western blot analysis was performed on whole cell extracts (50 ug lysate) of Panc-1 (Lane1), SH-SY5Y (Lane 2), SH-SY5Y treated with retinoic acid (10 uM/ 72 hours) (Lane 3), U-87MG (Lane 4), Jurkat (Lane 5), PC-3 (Lane 6) and PC-12 (Lane 7). The blots were probed with ABfinity™ Anti-Connexin 36 Recombinant Rabbit Oligoclonal Antibody (Product # 710663, 1-2 ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 ug/ml, 1:2500 dilution). A 72 kDa band corresponding to Connexin 36was observed across cell lines tested. Connexins are known to form dimers which justifies the 72 kDa molecular weight. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Rabbit|
|Host / Isotype||Goat / IgG|
|Immunogen||Recombinant full-length protein|
|Purification||Gravity column chromatography|
|Storage buffer||HRP Stabilizer, PBS|
|Storage Conditions||4° C, store in dark, DO NOT FREEZE!|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.05-1 µg/ml|
|Western Blot (WB)||0.05-1 µg/ml|
* 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.
The sensitivity and specificity of each lot is confirmed using ELISA.
Minimal cross-reactivity with mouse, rat, human, bovine, guinea pig and donkey IgG is observed.
Superclonal™ secondary antibodies are developed to be highly specific for both heavy and light chains of IgG. Utilizing the power of recombinant technology and multiple phenotypic screens, Superclonal™ antibodies are selected for enhanced specificity to the target species IgG. These recombinant secondary antibodies show similar performance as traditional polyclonal secondary antibodies in common immunoassay applications, with minimal cross-reactivity to other commonly-used species. In addition, their recombinant animal-free-origin helps ensures lot-to-lot consistency, reducing the need to optimize each lot in relevant assays.
Superclonal™ secondary antibodies are designed to allow scientists to achieve the accurate results they need, without some of the concerns that can impact traditional polyclonal secondary antibodies.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.