Product nameAnti-CstF-64 antibody
See all CstF-64 primary antibodies
DescriptionRabbit polyclonal to CstF-64
Tested applicationsSuitable for: IHC-P, WB, IP, ICC/IFmore details
Species reactivityReacts with: Human
Predicted to work with: Rabbit, Guinea pig, Cow, Pig, Chimpanzee, Rhesus monkey, Gorilla, Orangutan
Synthetic peptide corresponding to Human CstF-64 aa 375-425.
Database link: NP_001316.1
- HeLa whole cell lysate This antibody gave a positive result when used in the following formaldehyde/methanol fixed cell lines: MCF-7/HepG2. IHC-P: FFPE human testis tissue sections.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.09% Sodium azide
Constituents: 0.1% BSA, Tris buffered saline
Concentration information loading...
PurityImmunogen affinity purified
Purification notesab72297 was affinity purified using an epitope specific to CstF-64 immobilized on solid support.
Our Abpromise guarantee covers the use of ab72297 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|WB||1/2000 - 1/10000. Detects a band of approximately 68 kDa (predicted molecular weight: 61 kDa).|
|IP||Use at 2-5 µg/mg of lysate.|
|ICC/IF||Use a concentration of 5 µg/ml.|
FunctionOne of the multiple factors required for polyadenylation and 3'-end cleavage of mammalian pre-mRNAs. This subunit is directly involved in the binding to pre-mRNAs.
Sequence similaritiesContains 1 RRM (RNA recognition motif) domain.
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localizationNucleus. Localized with DDX1 in cleavage bodies.
- Information by UniProt
- betaCstF 64 variant 2 antibody
- CF-1 64 kDa subunit antibody
- CF1 64 kDa subunit antibody
All lanes : Anti-CstF-64 antibody (ab72297) at 0.04 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg
Lane 2 : HeLa whole cell lysate at 15 µg
Lane 3 : HeLa whole cell lysate at 5 µg
Developed using the ECL technique.
Predicted band size: 61 kDa
Observed band size: 68 kDa why is the actual band size different from the predicted?
Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 10 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling CstF-64 with ab72297 at 1/1000 (1µg/ml). Detection: DAB.
IHC image of CstF-64 staining in human testis formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab72297, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab72297 stained MCF-7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab72297 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HepG2 cells at 5ug/ml.
Detection of human CstF-64 by Immunoprecipitation using whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded), and ab72297 at 3 µg/mg lysate. Subsequent WB detection was performed using ab72297 at 0.1 µg/ml. Detection was by chemiluminescence with an exposure time of 3 seconds.
This product has been referenced in:
- Ko SH et al. Interference of DNAJB6/MRJ Isoform Switch by Morpholino Inhibits Replication of HIV-1 and RSV. Mol Ther Nucleic Acids 14:251-261 (2019). Read more (PubMed: 30641477) »
- Kim C et al. Regulation of post-translational protein arginine methylation during HeLa cell cycle. Biochim Biophys Acta 1800:977-85 (2010). WB ; Human . Read more (PubMed: 20541591) »