Overview

  • Product name

  • Description

    Rabbit polyclonal to XRN2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-P, IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chimpanzee, Rhesus monkey, Orangutan, Bat, Elephant
  • Immunogen

    Synthetic peptide corresponding to a region between residues 900 and 950 of human XRN2 (NP_036387.2)

  • Positive control

    • HeLa and 293T whole cell lysates. IHC-P: FFPE human breast adenocarcinoma.

Properties

Applications

Our Abpromise guarantee covers the use of ab72181 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
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP Use at 2-5 µg/mg of lysate.
WB 1/2000 - 1/10000. Detects a band of approximately 109 kDa (predicted molecular weight: 109 kDa).

Target

  • Relevance

    XRN2 possesses 5'->3' exoribonuclease activity. XRN2 shares similarity with the mouse Dhm1 and the yeast dhp1 gene. The function of the human gene has not yet been determined, however, it may promote the termination of transcription by RNA polymerase II. During transcription termination, cleavage at the polyadenylation site liberates a 5' fragment which is subsequently processed to form the mature mRNA and a 3' fragment which remains attached to the elongating polymerase. The processive degradation of this 3' fragment by this protein may promote termination of transcription.
  • Cellular localization

    Nucleus, nucleolus.
  • Database links

  • Alternative names

    • 5' 3' exoribonuclease 2 antibody
    • DHM1 like protein antibody
    • DHP protein antibody
    • XRN 2 antibody

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon carcinoma (left) and mouse teratoma (right) tissues labelling XRN2 with ab72181 at 1/200 (1µg/ml). Detection: DAB.
  • IHC image of XRN2 staining in human breast adenocarcinoma 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 ab72181, 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.

  • All lanes : Anti-XRN2 antibody (ab72181) 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
    Lane 4 : 293T whole cell lysate at 50 µg

    Predicted band size: 109 kDa
    Observed band size: 109 kDa
    Additional bands at: 170 kDa, 300 kDa, 70 kDa. We are unsure as to the identity of these extra bands.

  • Detection of Human XRN2 by Immunoprecipitation in Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded), using ab72181 at 3 µg/mg lysate for IP and at 1 µg/ml for subsequent Western blot detection.
  • All lanes : Anti-XRN2 antibody (ab72181) at 0.4 µg/ml

    Lane 1 : TCMK-1 whole cell lysate
    Lane 2 : 4T1 whole cell lysate
    Lane 3 : CT26 whole cell lysate

    Lysates/proteins at 50 µg per lane.

    Developed using the ECL technique.

    Predicted band size: 109 kDa


    Exposure time: 3 minutes
  • ICC/IF image of ab72181 stained MCF7 cells. The cells were 100% methanol fixed (5 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 (ab72181, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit 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.

References

This product has been referenced in:

  • Mersaoui SY  et al. Arginine methylation of the DDX5 helicase RGG/RG motif by PRMT5 regulates resolution of RNA:DNA hybrids. EMBO J 38:e100986 (2019). Read more (PubMed: 31267554) »
  • Charoy C  et al. Genetic specification of left-right asymmetry in the diaphragm muscles and their motor innervation. Elife 6:N/A (2017). WB ; Mouse . Read more (PubMed: 28639940) »
See all 10 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
ChIP
Sample
Mouse Cell lysate - whole cell (G1E)
Negative control
ChIP XRN2 on ALAS2 TSS and ChiP IgG
Specification
G1E
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 32 minute(s) and 0 second(s)
Specification of the cross-linking agent: EGS then Formaldehyde
Positive control
ChIP XRN2 from another vendor on TAL1 TTS

Abcam user community

Verified customer

Submitted Dec 22 2016

Question
Answer

The human epitope recognized by ab72181 is identical to the homologous rat epitope at 14 of 15 amino acids. In cases of even a one amino acid mismatch it can still be difficult to predict the chance for cross-species reactivity.  But since it is a polyclonal antibody the chances are good ab72181 will cross react with rat protein. We have a testing discount program you may be interested in. If you purchase a vial of ab72181 and test it in your rat samples and use our Abreview customer comment system to submit a short review of your experience, I can set up a discount code for you that will be good for a free antibody after we receive your Abreview. Please contact me if you are interested in this promotion or would like further information.

Read More
Application
Western blot
Sample
Mouse Tissue lysate - whole (Lung Tumors)
Loading amount
35 µg
Specification
Lung Tumors
Gel Running Conditions
Reduced Denaturing
Blocking step
Li-Cor Blocking Buffer as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Sep 17 2010

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