The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/200 - 1/250. (for 20 hrs - total 250 ug protein)
Use a concentration of 4 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use at an assay dependent concentration.
Use a concentration of 0.625 µg/ml. Detects a band of approximately 43 kDa (predicted molecular weight: 43 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
FunctionPlays an important role in caveolae formation and organization. Required for the sequestration of mobile caveolin into immobile caveolae. Termination of transcription by RNA polymerase I involves pausing of transcription by TTF1, and the dissociation of the transcription complex, releasing pre-rRNA and RNA polymerase I from the template. PTRF is required for dissociation of the ternary transcription complex.
Involvement in diseaseCongenital generalized lipodystrophy 4
Sequence similaritiesBelongs to the PTRF/SDPR family.
Post-translational modificationsPhosphorylated. Present in active and inactive forms. Changes in phosphorylation pattern may alter activity. Five truncated forms are found in the caveolae. These are thought to be the result of proteolysis and may be phosphorylation-dependent.
Cellular localizationMembrane > caveola. Cell membrane. Microsome. Endoplasmic reticulum. Cytoplasm > cytosol. Mitochondrion. Nucleus. Found at the surface of the caveolae. Also found in the plasma membrane, microsomal and cytosolic fractions and at a low level in the mitochondrial and nuclear fractions. Translocates to the cytoplasm from the caveolae upon insulin stimulation. CAV1 is necessary to recruit it to the cell membrane (By similarity). Co-localizes with CAV1 in lipid rafts in adipocytes.
ab48824 (4µg/ml) staining PTRF in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear and cytoplasmic staining. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
References for Anti-PTRF antibody (ab48824)
This product has been referenced in:
Shvets E et al. Dynamic caveolae exclude bulk membrane proteins and are required for sorting of excess glycosphingolipids. Nat Commun6:6867 (2015).
Read more (PubMed: 25897946) »
Swärd K et al. Arterial dysfunction but maintained systemic blood pressure in cavin-1-deficient mice. PLoS One9:e92428 (2014).
Read more (PubMed: 24658465) »