Recombinant
RabMAb

Recombinant Anti-IL-26 antibody [EPR22268-141] (ab224198)

Overview

  • Product name

    Anti-IL-26 antibody [EPR22268-141]
    See all IL-26 primary antibodies
  • Description

    Rabbit monoclonal [EPR22268-141] to IL-26
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human IL-26 aa 100 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9NPH9

  • Positive control

    • WB: THP-1 whole cell lysate; His-tagged human IL-26 recombinant protein (aa 22-171). IP: THP-1 treated with 80nM PMA (Phorbol-12-myristate-13-acetate) overnight, then 100ng/ml LPS (lipopolysaccharide) for 3h and add 300ng/ml BFA (Brefeldin A) for 3h, whole cell lysate. IHC-P: Human colon and cervical carcinoma tissue. ICC/IF: THP-1 cells treated with 80nM PMA (Phorbol-12-myristate-13-acetate) overnight, then 100ng/ml LPS (lipopolysaccharide) for 3h and add 300ng/ml BFA (Brefeldin A) for 3h.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22268-141
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab224198 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 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/30.
ICC/IF 1/100.
WB 1/1000. Detects a band of approximately 27 kDa (predicted molecular weight: 20 kDa).
  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    • Function

      May play a role in local mechanisms of mucosal immunity and seems to have a proinflammatory function. May play a role in inflammatory bowel disease. Activates STAT1 and STAT3, MAPK1/3 (ERK1/2), JUN and AKT. Induces expression of SOCS3, TNF-alpha and IL-8, secretion of IL-8 and IL-10 and surface expression of ICAM1. Decreases proliferation of intestinal epithelial cells. Is inhibited by heparin.
    • Tissue specificity

      Expressed in HVS transformed T-cells but not other T-cell lines or primary stimulated T-cells. Expressed in colonic T cells including Th17 inflammatory T cells; the expression is significantly increased in serum of patients with Crohn's disease (at protein level).
    • Sequence similarities

      Belongs to the IL-10 family.
    • Cellular localization

      Secreted.
    • Information by UniProt
    • Database links

    • Alternative names

      • AK155 antibody
      • IL 26 antibody
      • IL-26 antibody
      • IL26 antibody
      • IL26_HUMAN antibody
      • Interleukin 26 antibody
      • Interleukin-26 antibody
      • Protein AK155 antibody
      see all

    Images

    • All lanes : Anti-IL-26 antibody [EPR22268-141] (ab224198) at 1/1000 dilution

      Lane 1 : Untreated THP-1 (human monocytic leukemia cell line) whole cell lysate
      Lane 2 : THP-1 treated with 80nM PMA (Phorbol-12-myristate-13-acetate) for 24 hours, then 1µg/ml LPS (lipopolysaccharides) for 24 hours, whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 100000 mg/ml

      Predicted band size: 20 kDa
      Observed band size: 27 kDa
      why is the actual band size different from the predicted?


      Exposure time: 92 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The molecular weight observed is consistent with what has been described in the literature (PMID:14764663).

      This antibody reacts with an unidentifiable protein around 75 kDa.

    • Immunohistochemical analysis of paraffin-embedded human colon tissue labeling IL-26 with ab224198 at 1/1000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes of human colon (PMID: 18483078, 28852311) is observed. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. The section was incubated with ab224198 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized THP-1 (human monocytic leukemia cell line) cells labeling IL-26 with ab224198 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in THP-1 cell line treated with 80nM PMA (Phorbol-12-myristate-13-acetate) for 16h, then 100ng/ml LPS (lipopolysaccharide) for 3h and add 300ng/ml BFA (Brefeldin A) for 3h. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red). Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

      Cells were either untreated, or treated with 80nM PMA (Phorbol-12-myristate-13-acetate) for 16h, then 100ng/ml LPS (lipopolysaccharide) for 3h and add 300ng/ml BFA (Brefeldin A) for 3h.

    • IL-26 was immunoprecipitated from 0.35 mg of THP-1 (human monocytic leukemia cell line) (treated with 80nM PMA (Phorbol-12-myristate-13-acetate) overnight, then 100ng/ml LPS (lipopolysaccharide) for 3h and add 300ng/ml BFA (Brefeldin A) for 3h) whole cell lysate with ab224198 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224198 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/1000 dilution.

      Lane 1: THP-1 (treated as above) whole cell lysate 10 μg (Input).
      Lane 2: ab224198 IP in THP-1 (treated as above) whole cell lysate.
      Lane 3: : Rabbit monoclonal IgG (ab172730) instead of ab224198 in THP-1 (treated as above) whole cell lysate.

      Blocking/Dilution buffer: 5% NFDM/TBST.
      Exposure time: 3 mins.

    • Anti-IL-26 antibody [EPR22268-141] (ab224198) at 1/1000 dilution + His-tagged human IL-26 recombinant protein (aa 22-171), 20 ng

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 20 kDa


      Exposure time: 26 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemical analysis of paraffin-embedded human cervical carcinoma tissue labeling IL-26 with ab224198 at 1/1000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes of human cervical carcinoma (PMID: 18483078, 28852311) is observed. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. The section was incubated with ab224198 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    References

    ab224198 has not yet been referenced specifically in any publications.

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