Overview

  • Product name

  • Description

    Rabbit polyclonal to AGS3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide corresponding to Human AGS3 aa 100 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab90250)

  • Positive control

    • This antibody gave a positive signal in the following whole cell lysates: HEK293; HepG2. IHC-P: human colon adenocarcinoma FFPE tissue sections
  • General notes

     This product was previously labelled as RNase H2 subunit C

     

Properties

Applications

Our Abpromise guarantee covers the use of ab89726 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 before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 20 kDa (predicted molecular weight: 18 kDa).

Target

  • Relevance

    The RNase H2 complex is a heterotrimer composed of the catalytic subunit RNASEH2A and the non-catalytic subunits RNASEH2B and RNASEH2C (Ribonuclease H2 subunit C). Ribonuclease H2 subunit C is the non catalytic subunit of RNase H2, an endonuclease that specifically degrades the RNA of RNA:DNA hybrids. It participates in DNA replication, possibly by mediating the removal of lagging-strand Okazaki fragment RNA primers during DNA replication. Ribonuclease H2 subunit C also mediates the excision of single ribonucleotides from DNA:RNA duplexes.
  • Cellular localization

    Nuclear
  • Database links

  • Alternative names

    • AGS3 antibody
    • Aicardi-Goutieres syndrome 3 protein antibody
    • AYP1 antibody
    • Ribonuclease H2 subunit C antibody
    • Ribonuclease HI subunit C antibody
    • RNase H1 small subunit antibody
    • RNASEH2C antibody
    • RNH2C antibody
    see all

Images

  • All lanes : Anti-AGS3 antibody (ab89726) at 1 µg/ml

    Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell 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

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 18 kDa
    Observed band size: 20 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 45 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 12 minutes


    AGS3 contains a potential phosphorylation site (SwissProt) which may explain its migration at a higher molecular weight (20 kDa) than predicted (18 kDa).

  • IHC image of AGS3 staining in human colon adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab89726, 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-AGS3 antibody (ab89726) at 1 µg/ml

    All lanes : Mouse Pei1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 18 kDa
    Observed band size: 18 kDa


    Exposure time: 1 second


    Knockdown of Rnaseh2c using shRNA along with scramble control.

    Lane 1: scramble shRNA
    Lane 2: shRNA #1
    Lane 3: shRNA #2
    Lane 4: shRNA #3
    Lane 5: shRNA #4

    See Abreview

References

This product has been referenced in:

  • Shen T  et al. Antisense transcription regulates the expression of sense gene via alternative polyadenylation. Protein Cell 9:540-552 (2018). Read more (PubMed: 29273853) »
  • Uehara R  et al. Two RNase H2 Mutants with Differential rNMP Processing Activity Reveal a Threshold of Ribonucleotide Tolerance for Embryonic Development. Cell Rep 25:1135-1145.e5 (2018). Read more (PubMed: 30380406) »
See all 2 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (12%)
Sample
Mouse Cell lysate - whole cell (Pei1)
Specification
Pei1
Treatment
Knockdown of Rnaseh2c using shRNA along with scramble control
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Miss. Sarah Deasy

Verified customer

Submitted May 30 2014

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for WB and for human samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if you are also able to provide an image, including molecular weight markers, which would help us to assess the results.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.



Order Details
Antibody code:


Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


Amount of protein loaded


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to assess the results.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% TritonX100 in PBS
Specification
HeLa
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Jun 16 2010

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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