Western blot analysis was performed on nuclear enriched extracts (30 ug lysate) of K562 (Lane 1), Hep G2 (Lane 2), PC3 (Lane 3), and MCF-7 (Lane 4). The blots were probed with Anti-FKBP51 Rabbit Polyclonal Antibody (Product # PA1-020, 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). Approximately 51 and 45 kDa bands corresponding to FKBP51 were 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 with iBlot® 2 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||Dog, Hamster, Human, Mouse, Rat|
|Published species reactivity||Rat, Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to the residues C(394) Q K K A K E H N E R D R R(407) of human FKBP51.|
|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)||1-2 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 11 publications below|
PA1-020 detects FK506 binding protein 51 kDa (FKBP51) in canine, hamster, human, mouse and rat tissues.
PA1-020 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~51 kDa protein representing FKBP51 in HeLa cell lysate. By Western blot PA1-020 also detects FKBP51 in whole tissue extracts from rat kidney and rat and mouse testes.
The PA1-020 immunogen is a synthetic peptide corresponding to the residues C(394) Q K K A K E H N E R D R R(407) of human FKBP51. There is a single amino acid substitution (K396 to R396) in this domain between human and mouse FKBP51 protein. PA1-020 immunizing peptide (Cat. # PEP-078) is available for use in neutralization and control experiments.
Immunophilins are a family of soluble cytosolic receptors capable of binding to one of two major immunosuppressant agents - cyclosporin A (CsA) or FK506. Proteins that bind FK506 are termed FK506 Binding Proteins (FKBPs) and those that bind cyclosporin A are called cyclophilins (CyP). Both CyP:CsA and FKBP:FK506 complexes have been shown to inhibit calcineurin, a calcium and calmodulin dependent protein phosphatase which has been implicated as an important signaling enzyme in T-cell activation, providing a possible mechanism of immunosuppression by CsA and FK506. Immunophilins function as peptidyl prolyl cis-trans-isomerases (PPIase) whose activity is inhibited by their respective immunosuppressant compounds. As PPIase's, immunophilins accelerate folding of some proteins both in vivo and in vitro by catalyzing slow steps in the initial folding and rearrangement of proline containing proteins.
FK506 binding protein 51 kDa (FKBP51) is found associated with the progesterone receptor. This protein is also referred to as FKBP54 in the literature. This protein is predominantly expressed in murine T cells but in humans, it is abundantly expressed in numerous tissues at levels many times higher than FKBP12. The FKBP51 gene is known to be induced by glucocorticoids.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Pharmacological stimulation of hypoxia inducible factor-1¿ facilitates the corticosterone response to a mild acute stressor.
PA1-020 was used in western blot to determine if HIF-1 links glucocorticoids and oxidative stress
|Harrell CS,Rowson SA,Neigh GN||Neuroscience letters (600:75)||2015|
PHLPP1 splice variants differentially regulate AKT and PKC¿ signaling in hippocampal neurons: characterization of PHLPP proteins in the adult hippocampus.
PA1-020 was used in western blot to investigate the functions of different PHLPP splice variants in adult hippocampus
|Jackson TC,Verrier JD,Semple-Rowland S,Kumar A,Foster TC||Journal of neurochemistry (115:941)||2010|
The hsp90-FKBP52 complex links the mineralocorticoid receptor to motor proteins and persists bound to the receptor in early nuclear events.
PA1-020 was used in western blot to investigate the role of the heat shock protein 90-FKBP52 complex in the nucleus
|Galigniana MD,Erlejman AG,Monte M,Gomez-Sanchez C,Piwien-Pilipuk G||Molecular and cellular biology (30:1285)||2010|
Comparative analysis of calcineurin inhibition by complexes of immunosuppressive drugs with human FK506 binding proteins.
PA1-020 was used in western blot to study the inhibition of calcineurin by immunosuppressive drug-FK506 binding protein complexes
|Weiwad M,Edlich F,Kilka S,Erdmann F,Jarczowski F,Dorn M,Moutty MC,Fischer G||Biochemistry (45:15776)||2006|
Up-regulation of glucocorticoid-regulated genes in a mouse model of Rett syndrome.
PA1-020 was used in western blot to study the expression of glucocorticoid-regulated genes in Rett syndrome mouse model.
|Nuber UA,Kriaucionis S,Roloff TC,Guy J,Selfridge J,Steinhoff C,Schulz R,Lipkowitz B,Ropers HH,Holmes MC,Bird A||Human molecular genetics (14:2247)||2005|
Ethanol-responsive brain region expression networks: implications for behavioral responses to acute ethanol in DBA/2J versus C57BL/6J mice.
PA1-020 was used in western blot to investigate the gene expression profile in brain regions of the mesolimbic reward pathway of mouse strains DBA/2J and C57BL/6J .
|Kerns RT,Ravindranathan A,Hassan S,Cage MP,York T,Sikela JM,Williams RW,Miles MF||The Journal of neuroscience : the official journal of the Society for Neuroscience (25:2255)||2005|
Differential control of glucocorticoid receptor hormone-binding function by tetratricopeptide repeat (TPR) proteins and the immunosuppressive ligand FK506.
PA1-020 was used in western blot to investigate the regulatory effects of FK506 and TPR proteins on glucocorticoid receptor hormone-binding function
|Davies TH,Ning YM,Sánchez ER||Biochemistry (44:2030)||2005|
Association of immunophilins with mammalian TRPC channels.
PA1-020 was used in western blot to study the role of immunophilins in TRPC channel activation following receptor stimulation.
|Sinkins WG,Goel M,Estacion M,Schilling WP||The Journal of biological chemistry (279:34521)||2004|
Progesterone receptor deficient in chromatin binding has an altered cellular state.
PA1-020 was used in western blot to analyze the altered cellular processing by comparing the composition and distribution of tPR and sPR complexes
|Botos J,Xian W,Smith DF,Smith CL||The Journal of biological chemistry (279:15231)||2004|
A new first step in activation of steroid receptors: hormone-induced switching of FKBP51 and FKBP52 immunophilins.
PA1-020 was used in western blot to investigate the activation mechanism for glucocorticoid receptors.
|Davies TH,Ning YM,Sánchez ER||The Journal of biological chemistry (277:4597)||2002|
Tissue distribution and abundance of human FKBP51, and FK506-binding protein that can mediate calcineurin inhibition.
PA1-020 was used in western blot to study the expression of FK506-binding protein FKBP51 in tissues
|Baughman G,Wiederrecht GJ,Chang F,Martin MM,Bourgeois S||Biochemical and biophysical research communications (232:437)||1997|