Recombinant 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: WB, IP, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-TGN46 antibody [EPR12676]
See all TGN46 primary antibodies -
Description
Rabbit monoclonal [EPR12676] to TGN46 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, IHC-P, ICC/IFmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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/IF: Hep G2 and A549 cells.
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General notes
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR12676 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lysates
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Positive Controls
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Recombinant Protein
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 |
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WB |
1/1000 - 1/5000. Predicted molecular weight: 51 kDa.
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IP |
1/10 - 1/100.
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF |
Use a concentration of 1 µg/ml.
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Notes |
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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. |
ICC/IF
Use a concentration of 1 µg/ml. |
Target
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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
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Database links
- Entrez Gene: 10618 Human
- Omim: 603062 Human
- SwissProt: O43493 Human
- Unigene: 593382 Human
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Alternative names
- TGN 46 antibody
- TGN 51 antibody
- TGN38 antibody
see all
Images
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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.
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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.
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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. ab174280 at 1/1000 labeling TGN46 immunoprecipitated from Human fetal liver lysate using ab174280 at 1/10.
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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.
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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.
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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).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (1)
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