Overview

  • Product name
    Anti-Cleavage stimulation factor 2 antibody
    See all Cleavage stimulation factor 2 primary antibodies
  • Description
    Rabbit polyclonal to Cleavage stimulation factor 2
  • Tested applications
    Suitable for: IHC-P, WB, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Rabbit, Guinea pig, Cow, Pig, Chimpanzee, Rhesus monkey, Gorilla, Orangutan
  • Immunogen

    Synthetic peptide corresponding to a region between residues 375 and 425 of human Cleavage stimulation factor 2 (NP_001316.1)

  • Positive control
    • 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.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.09% Sodium Azide
    Constituents: 0.1% BSA, Tris buffered saline
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    ab72297 was affinity purified using an epitope specific to CSTF64 immobilized on solid support.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

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.

Application Abreviews Notes
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.

Target

  • Function
    One 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 similarities
    Contains 1 RRM (RNA recognition motif) domain.
  • Post-translational
    modifications
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization
    Nucleus. Localized with DDX1 in cleavage bodies.
  • Information by UniProt
  • Database links
  • Alternative names
    • betaCstF 64 variant 2 antibody
    • CF-1 64 kDa subunit antibody
    • CF1 64 kDa subunit antibody
    • Cleavage stimulation factor 64 kDa subunit antibody
    • Cleavage stimulation factor subunit 2 antibody
    • Cleavage stimulation factor, 3' pre RNA, subunit 2, 64kDa antibody
    • CSTF 2 antibody
    • CstF 64 antibody
    • CSTF 64 kDa subunit antibody
    • CstF-64 antibody
    • CSTF2 antibody
    • CSTF2_HUMAN antibody
    • MGC188397 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling Cleavage stimulation factor 2 with ab72297 at 1/1000 (1µg/ml). Detection: DAB.
  • IHC image of Cleavage stimulation factor 2 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.

  • All lanes : Anti-Cleavage stimulation factor 2 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
  • Detection of human Cleavage stimulation factor 2 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.

References

This product has been referenced in:
  • 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) »

See 1 Publication for this product

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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