Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Bovine, Monkey

Cited:

Human, Bovine, Primate - Macaca mulatta (Rhesus Macaque)

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunoblotting, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/ Immunofluorescence

Cited:

Western Blot, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all TGN46 Primary Antibodies »

Product Specifications

Immunogen

Partial recombinant protein made to an internal region of the human TGN46 protein (within residues 200-350). [Swiss-Prot: O43493]

Reactivity Notes

Monkey reactivity reported in scientific literature (PMID: 30135710). Bovine reactivity reported in scientific literature (PMID: 30135710).

Localization

Primarily in trans-Golgi network. Cycles between the trans-Golgi network and the cell surface returning via endosomes

Marker

TGN Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for TGN46 Antibody - BSA Free

Knockout Validated: TGN46 Antibody - BSA Free [NBP1-49643]

Western Blot: TGN46 Antibody - BSA Free [NBP1-49643]

Western Blot: TGN46 Antibody [NBP1-49643] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and TGN46 knockout (KO) HeLa cell line. PVDF membrane was probed with 1:1000 of Rabbit Anti-Human TGN46 Polyclonal Antibody (NBP1-49643) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). Specific band was detected for TGN46 at approximately 100 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.
Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

TGN46-Antibody-Immunocytochemistry-Immunofluorescence-NBP1-49643-img0021.jpg
Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

TGN46-Antibody-Immunocytochemistry-Immunofluorescence-NBP1-49643-img0022.jpg
Immunohistochemistry: TGN46 Antibody - BSA Free [NBP1-49643]

Immunohistochemistry: TGN46 Antibody - BSA Free [NBP1-49643]

Immunohistochemistry: TGN46 Antibody [NBP1-49643] - Analysis of TGOLN2 in human renal cell carcinoma.
Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

Immunocytochemistry/Immunofluorescence: TGN46 Antibody [NBP1-49643] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with TGN46 Antibody conjugated to Alexa Fluor 488 (NBP1-49643AF488) at 2 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Flow Cytometry: TGN46 Antibody - BSA Free [NBP1-49643]

Flow Cytometry: TGN46 Antibody - BSA Free [NBP1-49643]

Flow Cytometry: TGN46 Antibody [NBP1-49643] - An intracellular stain was performed on HepG2 cells with NBP1-49643C (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 650.
Western Blot: TGN46 AntibodyBSA Free [NBP1-49643]

Western Blot: TGN46 AntibodyBSA Free [NBP1-49643]

Western Blot: TGN46 Antibody [NBP1-49643] - Analysis of extracts from HeLa cells using TGN46 antibody (NBP1-49643, 1:100).
Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

Immunocytochemistry/Immunofluorescence: TGN46 Antibody [NBP1-49643] - HepG2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with TGN46 Antibody conjugated to Alexa Fluor 488 (NBP1-49643AF488) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

Immunocytochemistry/Immunofluorescence: TGN46 Antibody [NBP1-49643] - Analysis of A549 cells using TGN46 antibody (NBP1-49643, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

Immunocytochemistry/Immunofluorescence: TGN46 Antibody [NBP1-49643] - T98G glioblastoma cells probed for Golgi (Alexa Fluor 488 conjugated TGN-46 antibody, Green), tubulin (Alexa Fluor 594, Red), and nucleus (DAPI, blue). Image from the Alexa Fluor 488 version of this antibody. Image from verified customer review.
Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643]

Immunocytochemistry/Immunofluorescence: TGN46 Antibody [NBP1-49643] - HeLa cells were fixed in 4% paraformaldehyde for 10 min and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti- at 2 ug/ml for 60 minutes at room temperature and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective.
Flow (Intracellular): TGN46 Antibody - BSA Free [NBP1-49643]

Flow (Intracellular): TGN46 Antibody - BSA Free [NBP1-49643]

Flow (Intracellular): TGN46 Antibody [NBP1-49643] - An intracellular stain was performed on U-937 cells with NBP1-49643 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by rabbit IgG APC-conjugated secondary antibody (F0111, R&D Systems).
Flow (Intracellular): TGN46 Antibody - BSA Free [NBP1-49643]

Flow (Intracellular): TGN46 Antibody - BSA Free [NBP1-49643]

Flow (Intracellular): TGN46 Antibody [NBP1-49643] - An intracellular stain was performed on HepG2 cells with TGN46 Antibody NBP1-49643AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.
TGN46 Antibody - BSA Free

TGN46 in Caco-2 Human Cell Line.

TGN46 was detected in immersion fixed Caco-2 human colorectal adenocarcinoma cell line using Rabbit anti-TGN46 Affinity Purified Polyclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NBP1-49643AF488) (green) at 2 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.
TGN46 Antibody - BSA Free

TGN46 in HepG2 Human Cell Line.

TGN46 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Rabbit anti-TGN46 Affinity Purified Polyclonal Antibody conjugated to Biotin (Catalog # NBP1-49643B) at 2 µg/mL overnight at 4C. Cells were stained using Streptavidin conjugated to DyLight 550 (red) and counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.
TGN46 Antibody - BSA Free

TGN46 in HepG2 Human Cell Line -

TGN46 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Rabbit anti-TGN46 Affinity Purified Polyclonal Antibody conjugated to Alexa Fluor® 647 (Catalog # NBP1-49643AF647) (light blue) at 2 µg/mL overnight at 4C. Cells were counterstained with DAPI (dark blue). Cells were imaged using a 100X objective and digitally deconvolved.
TGN46 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643] -

Immunofluorescence microscopy of incoming virus particles, 20 minutes.(A, B) A549 cells were plated onto glass coverslips and the following day replaced with media containing NB-DNJ (200μM) or left untreated. Forty-eight hours later, cells were chilled to 4C on ice, then rVSV-SFTSV was bound by centrifugation (1200xg, 30’, 4C). Following centrifugation, media was replaced with pre-warmed media (37C) and the cells placed in a 37C incubator for 20 minutes before fixation in 2% paraformaldehyde for 10 minutes. Cells were then immunostained for viral antigen (anti-VSV M, red), cellular markers (green), and nuclei stained with DAPI (blue). Images are representative from at least 3 independent experiments. (A) A549 cells co-stained for rVSV-SFTSV (red) and early endosome marker EEA1 (green). (B) A549 cells co-stained for rVSV-SFTSV (red) and trans-Golgi marker TGN46 (green). Boxes indicate zoomed-in regions. Scale bar represents 5μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28388693), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TGN46 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643] -

Nonvesicular fraction of GII-NTPase significantly localizes to the ER or mitochondria in cells. (A) The expression plasmid encoding F-NTPase was transfected into A7 cells. After 24 h of transfection, cells were fixed and dual labeled with an anti-FLAG antibody (green) and a rabbit polyclonal anti-NTPase antibody (red). Colocalization of signals recognized by the anti-FLAG and the anti-NTPase antibodies was detected as yellow color. Scale bar, 50 μm. (B) A7 cells expressing F-NTPase were fixed and dual stained with an anti-FLAG antibody (green) and a set of antibodies or a fluorescent dye (MitoTracker [Mito]) specific for organelle markers (red). Colocalization of F-NTPase with the signals of organelle-specific markers are shown in yellow (arrows). The dashed boxes in each merged image were enlarged and are shown at the right. As noted, the nonvesicular areas of F-NTPase colocalize with the signals of PDI (an ER marker) or Mito (a mitochondrial probe) but not those of GM130 (a cis-Golgi marker), TGN46 (a trans-Golgi marker), and EEA1 (an endosomal marker). Scale bars, 10 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29212938), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TGN46 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643] -

Immunofluorescence microscopy of rVSV-SFTSV particles following UGCG knockdown.(A-D) U-2 OS cells were transfected with siRNAs targeting UGCG (siUGCG) or a non-targeting control (siNegCtrl) and plated onto glass coverslips. At 72 hours post-transfection cells were chilled to 4C on ice and rVSV-SFTSV was bound by centrifugation (1200xg, 30’, 4C). Following centrifugation, media was replaced with pre-warmed media (37C) and the cells placed in a 37C incubator for 20 or 40 minutes before fixation in 1% paraformaldehyde for 15 minutes. Cells were then immunostained for viral antigen (anti-VSV M, red), cellular markers (green), and nuclei stained with DAPI (blue). Images are representative from at least 3 independent experiments. (A,B) U-2 OS cells fixed after 20 minutes were co-stained for rVSV-SFTSV (red) and EEA1 (A) or TGN46 (B) (green). (C,D) U-2 OS cells fixed after 40 minutes and stained as above. Boxes indicate zoomed-in regions. Scale bar represents 5μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28388693), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TGN46 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: TGN46 Antibody - BSA Free [NBP1-49643] -

Immunofluorescence microscopy and quantification of incoming virus particles.(A, B) A549 cells prepared as described in Fig 8 were fixed 40 minutes post-warming and co-stained for rVSV-SFTSV (red) and the early endosome marker EEA1 (green) (A) or TGN46 (B). Cells were treated with NB-DNJ (bottom panels) or left untreated (top panels). Boxes indicate zoomed-in regions. Scale bar represents 5μm. (C, D) Quantitative image analysis was performed to measure the volume of discrete VSV M-stained puncta within z-stack images in untreated and NB-DNJ treated cells at both 20 and 40 minutes post-warming. Puncta were counted for at least 6 independent z-stacks per sample for both rVSV-SFTSV (C) and VSV (D) infected cells. (**** p<0.0001 using Welch’s one-tailed t-test). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28388693), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for TGN46 Antibody - BSA Free

Application
Recommended Usage

Immunoblotting

reported in scientific literature (PMID 25410859)

Immunocytochemistry/ Immunofluorescence

reported in scientific literature (PMID 31455601)

Immunohistochemistry

1:400

Immunohistochemistry-Paraffin

1:400

Western Blot

2ug/mL
Application Notes
Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

Reviewed Applications

Read 2 reviews rated 5 using NBP1-49643 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: TGN46

TGN46 (TGOLN2/trans-Golgi network protein 2) is a single-pass type I membrane protein localized in cell membrane as well as in trans-Golgi network (TGN) membrane, an important protein sorting station inside cells. It shuttles between TGN and cell surface returning via endosomes. TGN46-vesicle formation requires protein kinase D, a kinase needed for secretory carrier formation in cells. TGOLN2 gene has several alternatively spliced transcripts, of which isoform TGN46 is widely expressed whereas another isoform TGN51 is more abundant in fetal lung and kidney. TGN46 is a heavily glycosylated protein with largely unknown functions, however, it has been suspected to be involved in regulating membrane traffic to and from the TGN. TGN46 contains a signal peptide, lumenal domain, membrane-spanning domain, and cytoplasmic domain. Because the membrane spanning region and cytoplasmic tail contain the retention and retrieval signals, respectively, for localization in the TGN, TGN46 is considered a molecular marker for TGN. Recent expression studies have shown a connection between TGOLN2 and suicidal behavior wherein TGN46 was found to be up-regulated in suicide completers as compared to controls.

Alternate Names

TGN 46, TGN 48, TGN 51, TGN38, TGN-38, TGN38 homolog, TGN46, TGN-46, TGN48, TGN51, TGOLN 2, TGOLN2, trans golgi network 38, trans golgi network 46, Trans Golgi network integral membrane protein 2, Trans Golgi network integral membrane protein 2 [Precursor], Trans Golgi network protein (46 48 51kD isoforms), Trans golgi network protein 2, Trans Golgi network protein TGN51, Trans-Golgi network integral membrane protein 2, Trans-Golgi network protein 2, Trans-Golgi network protein TGN51, TTGN 2, TTGN2

Entrez Gene IDs

10618 (Human)

Gene Symbol

TGOLN2

UniProt

O43493 (Human)

Additional TGN46 Products

Product Documents for TGN46 Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for TGN46 Antibody - BSA Free

Manufactured by Genomic Antibody Technology™. GAT FAQs

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for TGN46 Antibody - BSA Free

Customer Reviews for TGN46 Antibody - BSA Free (2)

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Customer Images

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  • Name: Bryan Tinsley
    Application: Immunocytochemistry
    Sample Tested:
    Species: Human
    Verified Customer | Posted 06/12/2014
    IF Confocal analysis of A549 cells using TGN46 antibody (NBP1-49643, 1:5).
    TGN46 Antibody - BSA Free NBP1-49643
  • Name: Bryan Tinsley
    Application: Western Blot
    Sample Tested:
    Species: Human
    Verified Customer | Posted 06/12/2014
    Western blot analysis of extracts from HeLa cells using TGN46 antibody (NBP1-49643, 1:100).
    TGN46 Antibody - BSA Free NBP1-49643

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Protocols

View specific protocols for TGN46 Antibody - BSA Free (NBP1-49643):


Immunocytochemistry Protocol

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.



Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.

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FAQs

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