Recombinant
RabMAb

Recombinant Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224)

Overview

  • Product name

    Anti-Anterior Gradient 2 antibody [EPR20164-278]
    See all Anterior Gradient 2 primary antibodies
  • Description

    Rabbit monoclonal [EPR20164-278] to Anterior Gradient 2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Anterior Gradient 2 aa 1 to the C-terminus. The exact sequence is proprietary. Corresponds to the mature protein without the signal peptide.
    Database link: O95994

  • Positive control

    • WB: Human AGR2 recombinant protein; MCF7 and HT-29 whole cell lysates; Mouse, rat and human stomach lysates; Rat small intestine lysate. IHC-P: Human colon and ovary cancer tissues; Mouse and rat stomach tissues. ICC/IF: MCF7 cells. Flow Cyt: MCF7 cells.
  • 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

Applications

Our Abpromise guarantee covers the use of ab209224 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.
WB 1/1000. Detects a band of approximately 17 kDa (predicted molecular weight: 19 kDa).
ICC/IF 1/500.
Flow Cyt 1/70.

Target

  • Function

    Required for MUC2 post-transcriptional synthesis and secretion. May play a role in the production of mucus by intestinal cells (By similarity). Proto-oncogene that may play a role in cell migration, cell differentiation and cell growth.
  • Tissue specificity

    Expressed strongly in trachea, lung, stomach, colon, prostate and small intestine. Expressed weakly in pituitary gland, salivary gland, mammary gland, bladder, appendix, ovary, fetal lung, uterus, pancreas, kidney, fetal kidney, testis, placenta, thyroid gland and in estrogen receptor (ER)-positive breast cancer cell lines.
  • Sequence similarities

    Belongs to the AGR family.
  • Cellular localization

    Secreted. Endoplasmic reticulum.
  • Information by UniProt
  • Database links

  • Alternative names

    • AG 2 antibody
    • AG 2 protein antibody
    • AG-2 antibody
    • AG2 antibody
    • AG2 protein antibody
    • AGR 2 antibody
    • AGR2 antibody
    • AGR2_HUMAN antibody
    • Anterior gradient 2 homolog (Xenopus laevis) antibody
    • anterior gradient 2 homolog antibody
    • Anterior gradient 2, protein disulphide isomerase family member antibody
    • Anterior gradient 2, Xenopus, homolog of antibody
    • Anterior gradient homolog 2 antibody
    • Anterior gradient protein 2 homolog antibody
    • Epididymis secretory protein Li 116 antibody
    • GOB 4 antibody
    • GOB4 antibody
    • HAG 2 antibody
    • hAG-2 antibody
    • hAG2 antibody
    • HAG2/R antibody
    • HEL S 116 antibody
    • HPC 8 antibody
    • hPC8 antibody
    • PDIA17 antibody
    • Protein disulfide isomerase family A member 17 antibody
    • secreted cement gland homolog antibody
    • Secreted cement gland protein XAG 2 homolog antibody
    • Secreted cement gland protein XAG-2 homolog antibody
    • Secreted cement gland protein XAG2 homolog antibody
    • XAG 2 antibody
    • XAG2 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Anterior Gradient 2 with ab209224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on human colon is observed [PMID: 24794000].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • All lanes : Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224) at 1/2000 dilution

    Lane 1 : Human AGR2 recombinant protein
    Lane 2 : Human AGR3 recombinant protein

    Lysates/proteins at 0.01 µg per lane.

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

    Predicted band size: 19 kDa
    Observed band size: 17 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 10 seconds; Lane 2: 3 minutes.

    Human AGR2 recombinant protein contains aa21-175 with a His-tag, and human AGR3 recombinant protein contains aa21-166 with GST-tag.

  • All lanes : Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224) at 1/2000 dilution

    Lane 1 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
    Lane 2 : HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 19 kDa
    Observed band size: 17 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Negative control: HeLa (PMID: 16551856).

  • Lanes 1-3 : Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224) at 1/1000 dilution
    Lane 4 : Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224) at 1/2000 dilution

    Lane 1 : Mouse stomach lysate
    Lane 2 : Rat stomach lysate
    Lane 3 : Rat small intestine lysate
    Lane 4 : Human stomach lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lane 4 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 19 kDa
    Observed band size: 17 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-3: 3 minutes; Lane 4: 10 seconds.

  • Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue labeling Anterior Gradient 2 with ab209224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on human ovary cancer is observed [PMID: 22752467].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling Anterior Gradient 2 with ab209224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on mouse stomach is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling Anterior Gradient 2 with ab209224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on rat stomach is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling Anterior Gradient 2 with ab209224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Negative control: human pancreas [PMID: 20550709].

    Counter stained with Hematoxylin.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) and HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Anterior Gradient 2 with ab209224 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on MCF7 cell line.

    Negative control: HeLa (PMID: 16551856).

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed MCF7 (Human breast adenocarcinoma cell line) cells labeling Anterior Gradient 2 with ab209224 at 1/70 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Anterior Gradient 2 with ab209224 at 1/70 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

    Negative control: HeLa

References

This product has been referenced in:

  • Wang D  et al. Proteasome inhibition boosts autophagic degradation of ubiquitinated-AGR2 and enhances the antitumor efficiency of bevacizumab. Oncogene 38:3458-3474 (2019). Read more (PubMed: 30647455) »
See 1 Publication for this product

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