The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 2 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Substrate recognition component of a (SKP1-CUL1-F-box protein) E3 ubiquitin-protein ligase complex which mediates the ubiquitination and subsequent proteasomal degradation of target proteins. Probably recognizes and binds to phosphorylated target proteins during skeletal muscle atrophy. Recognizes TERF1.
Specifically expressed in cardiac and skeletal muscle.
All lanes : Anti-Fbx32 antibody (ab74023) at 2 µg/ml
Lane 1 : Human skeletal muscle tissue lysate - total protein (ab29330) Lane 2 : Skeletal Muscle (Mouse) Tissue Lysate Lane 3 : Skeletal Muscle (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 42 kDa Observed band size: 42 kDa Additional bands at: 90 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 15 minutes
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Fbx32 antibody (ab74023)This image is courtesy of an abreview submitted by Antibody Solutions Ltd.
ab74023 (1/2000) staining Fbx32 in paraffin-embedded Human heart tissue. Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval. The secondary antibody was goat anti rabbit conjugated to HRP. For further experimental details please refer to abreview.
Immunohistochemistry (Frozen sections) - Fbx32 antibody (ab74023)This image is courtesy of an Abreview submitted by Ruma Raha-Chowdhury
ab74023 staining Fbx32 in rat muscle tissue sections by Immunohistochemistry (Frozen section). Tissue was fixed with 4% PFA overnight. Samples were then blocked with 10% donkey serum for 1 hour at 24°C followed by incubation with ab74023 at a 1/200 dilution, for 24 hours at 4°C. A pig anti-rabbit Alexa Fluoro 488 conjugate was used as secondary antibody at a 1/1000 dilution. Nuclei were stained using the standard Hoestch method and are depicted in blue.
Immunocytochemistry/ Immunofluorescence - Anti-Fbx32 antibody (ab74023)This image is courtesy of an Abreview submitted by Ruma Raha-Chowdhury
ab74023 staining cultured rat astrocytes by ICC/IF. The cultured astrocytes were fixed with paraformaldehyde and blocked with 10% donkey serum for 30 minutes at 24°C. The cultured astrocytes were then stained with ab74023 at 1/500 in 0.3% TritonX with 0.1% PBS and 10% donkey serum for 4.5h at 24°C. An Alexa Fluro 488 donkey anti-rabbit polyclonal antibody at 1/1000 was used as the secondary antibody. Nuclei were stained with 1.43µM Hoechst and can be observed in blue. FBX32 expressed in the newly born astrocytes but does not co-localise with mature astrocytes.
Li F et al. Ginsenoside Rg1 prevents starvation-induced muscle protein degradation via regulation of AKT/mTOR/FoxO signaling in C2C12 myotubes. Exp Ther Med14:1241-1247 (2017).
Read more (PubMed: 28781621) »