Western blot analysis was performed on whole cell extracts (30 ug) of THP-1 (Lane 1), COLO 205(Lane 2), HCT 116 (Lane 3), Jurkat(Lane 4), HeLa(Lane 5), MCF7(Lane 6) and MDA-MB-231 (Lane 7).The blots were probed with Anti-AIP Rabbit Polyclonal Antibody (Product # PA1-514, 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 ~45 kDa band corresponding to AIP was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Full length, bacterially expressed mouse recombinant AIP.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Western Blot (WB)||2 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.
|Western Blot (WB)||See 1 publications below|
PA1-514 detects aryl hydrocarbon receptor interacting protein (AIP) from mouse samples.
PA1-514 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~38 kDa protein representing AIP from Hepa 1 SV40 cell lysate, as well as recombinant mouse AIP.
PA1-514 immunogen is full length, bacterially expressed mouse recombinant AIP.
Studies have shown that the unactivated aryl hydrocarbon receptor (AhR) exists in a heterotetrameric 9S core with hsp90 and the aryl hydrocarbon receptor interacting protein (AIP, also known as ARA9 or XAP-2). AIP is a 330 amino acid protein that resembles the 52 kDa FKBP52 protein sequence, but does not share the same affinity for the hsp90-glucocorticoid receptor complex.
It has been demonstrated through studies that AIP helps stabilize the AIP-hsp90-AhR complex and protects the AhR receptor from ubiquitination under heat stress conditions. These studies also suggest that AIP plays an important role in regulating the rate of AhR turnover by delaying AhR nuclear translocation after ligand binding. Immunopreciatation experiments have also shown that the N-terminus of AIP is required for the stability of the dioxin receptor-hsp90-AIP complex and the C-terminus is needed for direct contact and binding to hsp90.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The cholinergic and non-cholinergic effects of organophosphates and oximes in cultured human myoblasts.
PA1-514 was used in western blot to study the effect of organophosphates and oximes on cholinergic and non-cholinergic processes in human myoblasts
|Katalini¿ M,Mi¿ K,Pirkmajer S,Grubi¿ Z,Kovarik Z,Mar¿ T||Chemico-biological interactions (203:144)||2013|