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Also for FBXO4 (NM_134099)
|This whole rabbit serum was prepared by repeated immunizations with a synthetic peptide derived from sequences unique to the N-terminus of FBX4 and conserved between the human and murine proteins (see link below for the full-length sequence of the mouse gene product).|
||ELISA: 1:5,000 - 1:25,000, WB: 1:500 – 1:3000
|0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2|
|Skp/Cullin/F-box (SCF) complexes are part of a large family of E3 ubiquitin ligases in which F-box proteins determine substrate specificity. Substrate selection by F-box proteins may also be aided by accessory cofactors. The F-box only protein 4, FBX4, interacts via its F-box with SKP1 and CUL1 to form SCFFBX4 ubiquitin ligase. The activity of the SCFFBX4 complex can be regulated by aB crystallin, a small heat shock protein which serves as a specificity cofactor for FBX4 dependent ubiquitination. FBX4 together with aB crystallin (SCFFBX4-aB crystallin) promotes the ubiquitination of cyclin D1 phosphorylated at Thr-286. In addition, the SCFFBX4-aB crystallin ubiquitin ligase has been implicated in the ubiquitination of unknown substrates in desmin-related myopathy. SCFFBX4, in the absence of aB crystallin, also ubiquitinates the telomeric protein PIN2/TRF1.
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Whole cell extracts prepared from U2OS cells, or insect Sf9 cells expressing Flag-FBX4, were resolved by SDS-PAGE and transferred to nitrocellulose. FBX4 was detected using Rockland’s anti-FBX4 at a 1:500 dilution, in TBS buffer containing 0.1% Tween-20, followed by peroxidase conjugated anti-rabbit IgG. Personal Communication. A, Diehl, Univ. of Pennsylvania, Philadelphia, PA.
Immunocytochemistry staining of NIH-3T3 cells with Rockland’s anti-FBX4. Cells were fixed in methanol/acetone followed by incubation with anti-FBX4 at a 1:300 dilution. Personal Communication. A, Diehl, Univ. of Pennsylvania, Philadelphia,