Anti-PADI2 / PAD2 antibody (ab16478)

Reviews (2)Q&A (1)References (13)
Western blot - Anti-PADI2 / PAD2 antibody (ab16478)
  • Immunocytochemistry/ Immunofluorescence - Anti-PADI2 / PAD2 antibody (ab16478)

Key features and details

  • Rabbit polyclonal to PADI2 / PAD2
  • Suitable for: ICC/IF, WB
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-PADI2 / PAD2 antibody
    See all PADI2 / PAD2 primary antibodies

  • Description

    Rabbit polyclonal to PADI2 / PAD2

  • Host species

    Rabbit

  • Specificity

    ab16478 recognises a specific 75kDa band corresponding to PADI2, which is specifically blocked using the immunizing peptide in human colon, skeletal muscle and kidney lysates. There is a non-specific 18kDa band present in skeletal muscle lysates, which is attributed to cross-reactivity of the PADI2 antibody

  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide corresponding to Human PADI2/ PAD2 aa 100-200 (internal sequence) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab17091)

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab16478 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
WB
Human
All applications
Rat
Application Abreviews Notes
ICC/IF
Use a concentration of 5 µg/ml.
WB (1)
Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 76 kDa).
Notes
ICC/IF
Use a concentration of 5 µg/ml.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 76 kDa).

Target

  • Function

    Catalyzes the deimination of arginine residues of proteins.

  • Sequence similarities

    Belongs to the protein arginine deiminase family.

  • Cellular localization

    Cytoplasm.
  • Information by UniProt

  • Database links

    • Alternative names

      • KIAA0994 antibody
      • OTTHUMP00000044625 antibody
      • PAD 2 antibody
      • PAD H19 antibody
      • PAD-H19 antibody
      • PAD2 antibody
      • PADI 2 antibody
      • Padi2 antibody
      • PADI2 protein antibody
      • PADI2_HUMAN antibody
      • PDI 2 antibody
      • PDI2 antibody
      • Peptidlyarginine deiminase type II antibody
      • Peptidyl arginine deiminase II antibody
      • Peptidyl arginine deiminase type II antibody
      • Peptidylarginine deiminase II antibody
      • Protein arginine deiminase antibody
      • Protein arginine deiminase type 2 antibody
      • Protein arginine deiminase type II antibody
      • Protein-arginine deiminase type II antibody
      • Protein-arginine deiminase type-2 antibody
      see all

    Images

    • Western blot - Anti-PADI2 / PAD2 antibody (ab16478)
      Western blot - Anti-PADI2 / PAD2 antibody (ab16478)
      Anti-PADI2 / PAD2 antibody (ab16478) at 1 µg/ml + Human kidney tissue lysate - total protein (ab30203) at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 76 kDa
      Observed band size: 75 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 34 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 20 minutes


      This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab16478 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution. The 75-kDa band observed is consistent with what has been described in the literature (PMID:18668562; 20668670; 16723463).

       
    • Immunocytochemistry/ Immunofluorescence - Anti-PADI2 / PAD2 antibody (ab16478)
      Immunocytochemistry/ Immunofluorescence - Anti-PADI2 / PAD2 antibody (ab16478)
      ICC/IF image of ab16478 stained human Hek293 cells. The cells were PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab16478, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

    References (13)

    Publishing research using ab16478? Please let us know so that we can cite the reference in this datasheet.

    ab16478 has been referenced in 13 publications.

    • Casanova V  et al. Citrullination Alters the Antiviral and Immunomodulatory Activities of the Human Cathelicidin LL-37 During Rhinovirus Infection. Front Immunol 11:85 (2020). PubMed: 32117246
    • Gao BS  et al. Peptidyl Arginine Deiminase, Type II (PADI2) Is Involved in Urothelial Bladder Cancer. Pathol Oncol Res N/A:N/A (2019). PubMed: 31267364
    • García-Berrocoso T  et al. Single Cell Immuno-Laser Microdissection Coupled to Label-Free Proteomics to Reveal the Proteotypes of Human Brain Cells After Ischemia. Mol Cell Proteomics 17:175-189 (2018). PubMed: 29133510
    • Zhou Y  et al. Spontaneous Secretion of the Citrullination Enzyme PAD2 and Cell Surface Exposure of PAD4 by Neutrophils. Front Immunol 8:1200 (2017). PubMed: 28993780
    • McNee G  et al. Citrullination of histone H3 drives IL-6 production by bone marrow mesenchymal stem cells in MGUS and multiple myeloma. Leukemia 31:373-381 (2017). PubMed: 27400413
    View all Publications for this product

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Question

    Sehr geehrte Damen und Herren,

    habe den Antikörper anti-PAD2 (ab16478) im Western Blott eingesetzt und war etwas verwundert weil der Antikörper sehr spezifisch eine Bande bei ca. 75 kDa erkennt (im Kontroll Lysat Hek293 cells). Im Daten Sheet steht drin, dass eigentlich eine Bande von 45 kDa erkannt werden soll. Mir ist auch der kleine Vermerk aufgefallen, in dem steht das die Größe von PAD2 noch nicht geklärt ist. In der Literatur findet man aber eigentlich nur die Info dass PAD2 75 kDa groß ist.

    Meine Frage: Wie kann das denn sein dass der Antikörper das kleinere erkennen soll? Bzw. gibt es einen Literaturnachweis für die kleinere Version? Die im Sheet angegebene ist nicht sehr befriedigend.

    Vielen Dank schon im Vorraus
    und freundliche Grüße

    Read More
    Answer

    Vielen Dank für Ihre Email.

    Leider weiss ich auch nicht genau, was der Unterschied zwischen diesen beiden Banden ist. In UniProt hat es zwei Sequenzen - die annotierte lange Form (http://www.uniprot.org/uniprot/Q9Y2J8) und die noch un- annotierte kurze Form (http://www.uniprot.org/uniprot/Q96DA7).

    Da Sie die HEK293 Zellen benutzen, und wir Nierenlysat benutzt haben, kann ich Ihnen vorschlagen, ein gratis Röhrchen von Nierenlysat als positive Kontrolle zu senden. Dann könnten Sie gleich die beiden Proben vergleichen.

    Es tut mir leid, Ihnen nicht andersweitig helfen zu können. Es scheint mir, dass die Proteinbiologie von PADI2 noch nicht genau erforscht ist.

    Bitte lassen Sie mich wissen, ob Sie dies akzeptieren können und wollen.

    Read More

    Flow Cytometry abreview for Anti-PADI2 / PAD2 antibody

     Excellent
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (Peripheral blood mononuclear cells)
    Specification
    Peripheral blood mononuclear cells
    Preparation
    Cell harvesting/tissue preparation method: Normal human peripheral blood monocytes cultured with M-CSF. Cells were harvested on day 3, non-specific binding was blocked with human Ig, staining was performed on permeabilized cells
    Sample buffer: flow cytometery staining buffer (PBS 0.1% sodium azide 1% BSA) after permeabilization
    Fixation
    Caltag Fix Perm Kit
    Permeabilization
    Yes - Caltag Fix Perm Kit
    Gating Strategy
    Myeloid gate
    Read More

    Dr. Frances Santiago-Schwarz

    Verified customer

    Submitted Jul 01 2009

    Western blot abreview for Anti-PADI2 / PAD2 antibody

     Below Average
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (THP-1, MRC-5, human macrophages)
    Loading amount
    1e+006 cells
    Specification
    THP-1, MRC-5, human macrophages
    Blocking step
    Other as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 10 2006

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