Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] - BSA and Azide free (AB283702)

This data was developed using ab283678, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized C2C12 cells labelling TGN38 with ab283678 at 1/50 (11.38 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing co-locolization positive staining with Golgi marker in C2C12 cells is observed. ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain tubulin at 1/200 5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] - BSA and Azide free (AB283702)

This data was developed using ab283678, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling TGN38 with ab283678 at 1/50 (11.38 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing co-locolization positive staining with Golgi marker in NIH/3T3 cells is observed. ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain tubulin at 1/200 5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] - BSA and Azide free (AB283702)

This data was developed using ab283678, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling TGN38 with ab283678 at 1/2000 (0.285 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Granular cytoplasmic staining in mouse cerebrum. The section was incubated with ab283678 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] - BSA and Azide free (AB283702)

This data was developed using ab283678, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling TGN38 with ab283678 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat F(ab')2 Anti-Rabbit IgG (DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] - BSA and Azide free (AB283702)

This data was developed using ab283678, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labelling TGN38 with ab283678 at 1/2000 (0.285 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Granular cytoplasmic staining in mouse liver. The section was incubated with ab283678 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] - BSA and Azide free (AB283702)

This data was developed using ab283678, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labelling TGN38 with ab283678 at 1/2000 (0.285 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Granular cytoplasmic staining in mouse kidney. The section was incubated with ab283678 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] - BSA and Azide free (AB283702)

This data was developed using ab283678, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C2C12 (Mouse myoblasts myoblast) cells labelling TGN38 with ab283678 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat F(ab')2 Anti-Rabbit IgG (DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] - BSA and Azide free (AB283702)

This data was developed using ab283678, the same antibody clone in a different buffer formulation.

TGN38 was immunoprecipitated from 0.35 mg C2C12 (mouse myoblasts myoblast), whole cell lysate 10 ug with ab283678 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283678 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : C2C12 (mouse myoblasts myoblast), whole cell lysate 10 ug

Lane 2 : ab283678 IP in C2C12 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283678 in C2C12 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 49 seconds

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Immunoprecipitation - Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] (<a href='/en-us/products/primary-antibodies/trans-golgi-network-integral-membrane-protein-1-antibody-epr24831-36-ab283678'>ab283678</a>)

Predicted band size: 38 kDa

Observed band size: 75 kDa

false

• WB

Lab

Western blot - Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] - BSA and Azide free (AB283702)

This data was developed using ab283678, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

This antibody has cross-reaction with mouse TGN38B.

The recombinant proteins TGN38B and TGN38 were made in-house.

The sample loaded onto lane 1 was purified TGN38B recombinant protein expressed from an E.coli expression system.

The sample loaded onto lanes 2 was purified TGN38 recombinant protein expressed from a mammalian - HEK-293 expression system.

Exposure time : 81 seconds

All lanes:

Western blot - Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] (<a href='/en-us/products/primary-antibodies/trans-golgi-network-integral-membrane-protein-1-antibody-epr24831-36-ab283678'>ab283678</a>) at 1/1000 dilution

Lane 1:

His-tagged mouse TGN38B recombinant protein at 0.01 µg

Lane 2:

LIF-tagged mouse TGN38 recombinant protein at 0.01 µg

Secondary

Lane 1:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Lane 2:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 38 kDa

Observed band size: 50 kDa,65-100 kDa

false

• WB

Lab

Western blot - Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] - BSA and Azide free (AB283702)

This data was developed using ab283678, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Lane 1 of this blot was developed using a higher sensitivity ECL substrate.

The expression profiles/observed MW are consistent with what has been described in the literature (PMID : 12494998).

Exposure time : 3 minutes

All lanes:

Western blot - Anti-Trans-Golgi network integral membrane protein 1 antibody [EPR24831-36] (<a href='/en-us/products/primary-antibodies/trans-golgi-network-integral-membrane-protein-1-antibody-epr24831-36-ab283678'>ab283678</a>) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Lane 2:

C2C12 (mouse myoblasts myoblast), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 38 kDa

Observed band size: 85 kDa

false