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Anti-PARN Antibody Picoband™ (monoclonal, 11D8)

     
  • 1 - Anti-PARN Antibody Picoband™ (monoclonal, 11D8) ABO14826
    Figure 1. Western blot analysis of PARN using anti-PARN antibody (M01501).
    Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
    Lane 1: human Hela whole cell lysates,
    Lane 2: human 293T whole cell lysates,
    Lane 3: monkey COS-7 whole cell lysates,
    Lane 4: human CACO-2 whole cell lysates,
    Lane 5: rat kidney tissue lysates,
    Lane 6: rat C6 whole cell lysates,
    Lane 7: mouse NIH/3T3 whole cell lysates.
    After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-PARN antigen affinity purified monoclonal antibody (Catalog # M01501) at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for PARN at approximately 78KD. The expected band size for PARN is at 73KD.
  • 2 - Anti-PARN Antibody Picoband™ (monoclonal, 11D8) ABO14826
    Figure 2. IHC analysis of PARN using anti PARN antibody (M01501).
    PARN was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-PARN Antibody (M01501) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
  • 3 - Anti-PARN Antibody Picoband™ (monoclonal, 11D8) ABO14826
    Figure 3. IF analysis of PARN using anti-PARN antibody (M01501).
    PARN was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL mouse anti-PARN Antibody (M01501) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
  • 3 - Anti-PARN Antibody Picoband™ (monoclonal, 11D8) ABO14826
    Figure 4. IF analysis of PARN using anti-PARN antibody (M01501).
    PARN was detected in immunocytochemical section of U251 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL mouse anti-PARN Antibody (M01501) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
  • 1 - Anti-PARN Antibody Picoband™ (monoclonal, 11D8) ABO14826
    Figure 5. Flow Cytometry analysis of A549 cells using anti-PARN antibody (M01501).
    Overlay histogram showing A549 cells stained with M01501 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-PARN Antibody (M01501, 1 µg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
  • 1 - Anti-PARN Antibody Picoband™ (monoclonal, 11D8) ABO14826
    Figure 6. Flow Cytometry analysis of A431 cells using anti-PARN antibody (M01501).
    Overlay histogram showing A431 cells stained with M01501 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-PARN Antibody (M01501, 1 µg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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Product Information
Application
  • Applications Legend:
  • E=ELISA
  • WB=Western Blotting
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin)
  • IP=Immunoprecipitation
  • IF=Immunofluorescence
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • FC=Flow Cytometry
  • DB=Dot Blot
WB, IHC, IF, ICC, FC
Primary Accession O95453
Host Mouse
Isotype Mouse IgG2b
Reactivity Rat, Human, Mouse, Monkey
Clonality Monoclonal
Format Lyophilized
Description Anti-PARN Antibody Picoband™ (monoclonal, 11D8) . Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Monkey, Mouse, Rat.
Reconstitution Add 0.2ml of distilled water will yield a concentration of 500 µg/ml.
Additional Information
Gene ID 5073
Other Names Poly(A)-specific ribonuclease PARN, 3.1.13.4, Deadenylating nuclease, Deadenylation nuclease, Polyadenylate-specific ribonuclease, PARN, DAN
Calculated MW 73451 Da
Application Details Western blot, 0.1-0.5 µg/ml
Immunohistochemistry (Paraffin-embedded Section), 0.5-1 µg/ml
Immunocytochemistry/Immunofluorescence, 2 µg/ml
Flow Cytometry, 1-3 µg/1x10^6 cells
Subcellular Localization Nucleus. Nucleolus. Cytoplasm.
Tissue Specificity Ubiquitous.
Source Eukaryota
Contents Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clone Names Clone: 11D8
Immunogen E. coli-derived human PARN recombinant protein (Position: M1-Y301).
Cross Reactivity No cross-reactivity with other proteins.
Storage Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name PARN
Synonyms DAN
Function 3'-exoribonuclease that has a preference for poly(A) tails of mRNAs, thereby efficiently degrading poly(A) tails. Exonucleolytic degradation of the poly(A) tail is often the first step in the decay of eukaryotic mRNAs and is also used to silence certain maternal mRNAs translationally during oocyte maturation and early embryonic development. Interacts with both the 3'-end poly(A) tail and the 5'-end cap structure during degradation, the interaction with the cap structure being required for an efficient degradation of poly(A) tails. Involved in nonsense-mediated mRNA decay, a critical process of selective degradation of mRNAs that contain premature stop codons. Also involved in degradation of inherently unstable mRNAs that contain AU- rich elements (AREs) in their 3'-UTR, possibly via its interaction with KHSRP. Probably mediates the removal of poly(A) tails of AREs mRNAs, which constitutes the first step of destabilization (PubMed:10882133, PubMed:11359775, PubMed:12748283, PubMed:15175153, PubMed:9736620). Also able to recognize and trim poly(A) tails of microRNAs such as MIR21 and H/ACA box snoRNAs (small nucleolar RNAs) leading to microRNAs degradation or snoRNA increased stability (PubMed:22442037, PubMed:25049417).
Cellular Location Nucleus. Cytoplasm. Nucleus, nucleolus. Note=Some nuclear fraction is nucleolar
Tissue Location Ubiquitous.
Research Areas

BACKGROUND

Poly (A)-specific ribonuclease (PARN), also known as polyadenylate-specific ribonuclease or deadenylating nuclease (DAN), is an enzyme that in humans is encoded by the PARN gene. The protein encoded by this gene is a 3'-exoribonuclease, with similarity to the RNase D family of 3'-exonucleases. It prefers poly (A) as the substrate, hence, efficiently degrades poly (A) tails of mRNAs. Exonucleolytic degradation of the poly (A) tail is often the first step in the decay of eukaryotic mRNAs. This protein is also involved in silencing of certain maternal mRNAs during oocyte maturation and early embryonic development, as well as in nonsense-mediated decay (NMD) of mRNAs that contain premature stop codons.

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