Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-TGN46 antibody [EPR12676] (ab174280)

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Western blot - Anti-TGN46 antibody [EPR12676] (ab174280)
  • Western blot - Anti-TGN46 antibody [EPR12676] (ab174280)
  • Immunocytochemistry - Anti-TGN46 antibody [EPR12676] (ab174280)
  • Immunoprecipitation - Anti-TGN46 antibody [EPR12676] (ab174280)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TGN46 antibody [EPR12676] (ab174280)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TGN46 antibody [EPR12676] (ab174280)
  • Immunocytochemistry - Anti-TGN46 antibody [EPR12676] (ab174280)
  • Anti-TGN46 antibody [EPR12676] (ab174280)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR12676] to TGN46
  • Suitable for: ICC, WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-TGN46 antibody [EPR12676]
    See all TGN46 primary antibodies

  • Description

    Rabbit monoclonal [EPR12676] to TGN46

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC, WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A549, HepG2 and HeLa lysates (HeLa shows to be a low expressor). IHC-P: Human brain and kidney tissues. IP: Immunoprecipitation pellet from fetal liver lysate. ICC: Hep G2 and A549 cells.

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

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

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR12676

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab174280 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
ICC
Use a concentration of 1 µg/ml.
WB
1/1000 - 1/5000. Predicted molecular weight: 51 kDa.
IP
1/10 - 1/100.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
ICC
Use a concentration of 1 µg/ml.
WB
1/1000 - 1/5000. Predicted molecular weight: 51 kDa.
IP
1/10 - 1/100.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt.

    • Target

    • Function

      May be involved in regulating membrane traffic to and from trans-Golgi network.

    • Tissue specificity

      Isoform TGN46 is widely expressed. Isoform TGN51 is more abundant in fetal lung and kidney. Isoform TGN48 is barely expressed in embryonic kidney and promyelocytic cells.

    • Cellular localization

      Cell membrane. Golgi apparatus > trans-Golgi network membrane. Primarily in trans-Golgi network. Cycles between the trans-Golgi network and the cell surface returning via endosomes.
    • Information by UniProt

    • Database links

      • Alternative names

        • TGN 46 antibody
        • TGN 51 antibody
        • TGN38 antibody
        • TGN38 homolog antibody
        • TGN38B antibody
        • TGN46 antibody
        • TGN48 antibody
        • TGN51 antibody
        • TGOLN2 antibody
        • TGON2_HUMAN antibody
        • Trans Golgi network integral membrane protein 2 antibody
        • Trans golgi network protein 2 antibody
        • Trans Golgi network protein TGN51 antibody
        • Trans-Golgi network integral membrane protein 2 antibody
        • Trans-Golgi network protein TGN51 antibody
        • TTGN2 antibody
        see all

      Images

      • Western blot - Anti-TGN46 antibody [EPR12676] (ab174280)
        Western blot - Anti-TGN46 antibody [EPR12676] (ab174280)
        All lanes : Anti-TGN46 antibody [EPR12676] (ab174280) at 1/1000 dilution

        Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
        Lane 2 : TGOLN2 knockout A549 (Human lung carcinoma cell line) whole cell lysate
        Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
        Lane 4 : Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

        Lysates/proteins at 20 µg per lane.

        Performed under reducing conditions.

        Predicted band size: 51 kDa
        Observed band size: 80 kDa why is the actual band size different from the predicted?



        Lanes 1 - 4: Merged signal (red and green). Green - ab174280 observed at 80 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

        ab174280 was shown to react with TGN46 in wild-type A549 cells in Western blot with loss of signal observed in TGOLN2 knockout cell line ab269510 (knockout cell lysate ab269672). Wild-type A549 and TGOLN2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab174280 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

      • Western blot - Anti-TGN46 antibody [EPR12676] (ab174280)
        Western blot - Anti-TGN46 antibody [EPR12676] (ab174280)
        All lanes : Anti-TGN46 antibody [EPR12676] (ab174280) at 1/1000 dilution

        Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
        Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

        Predicted band size: 51 kDa
        Observed band size: 75-100 kDa why is the actual band size different from the predicted?



        Blocking buffer: 5% NFDM/TBST.

        Exposure: 10 seconds, 100 seconds.

      • Immunocytochemistry - Anti-TGN46 antibody [EPR12676] (ab174280)
        Immunocytochemistry - Anti-TGN46 antibody [EPR12676] (ab174280)
        ab174280 staining TGN46 in wild-type A549 cells (top panel) and TGOLN2 knockout A549 cells (bottom panel) (ab269510). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab174280 at 1μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
        Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
      • Immunoprecipitation - Anti-TGN46 antibody [EPR12676] (ab174280)
        Immunoprecipitation - Anti-TGN46 antibody [EPR12676] (ab174280)

        Immunoprecipitation. ab174280 at 1/1000 labeling TGN46 immunoprecipitated from Human fetal liver lysate using ab174280 at 1/10.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TGN46 antibody [EPR12676] (ab174280)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TGN46 antibody [EPR12676] (ab174280)

        Immunohistochemical analysis of paraffin embedded Human brain tissue labeling TGN46 with ab174280 at 1/50.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TGN46 antibody [EPR12676] (ab174280)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TGN46 antibody [EPR12676] (ab174280)

        Immunohistochemical analysis of paraffin embedded Human kidney tissue labeling TGN46 with ab174280 at 1/50.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunocytochemistry - Anti-TGN46 antibody [EPR12676] (ab174280)
        Immunocytochemistry - Anti-TGN46 antibody [EPR12676] (ab174280)

        ab174280 staining TGN46 in HepG2 cells (top panel - positive control) and HeLa cells (bottom panel - negative control, low TGN46 expression). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab174280 at 1μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
        Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

      • Anti-TGN46 antibody [EPR12676] (ab174280)
        Anti-TGN46 antibody [EPR12676] (ab174280)

      References (1)

      Publishing research using ab174280? Please let us know so that we can cite the reference in this datasheet.

      ab174280 has been referenced in 1 publication.

      • Han J  et al. HSP90 inhibition targets autophagy and induces a CASP9-dependent resistance mechanism in NSCLC. Autophagy 14:958-971 (2018). PubMed: 29561705

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