Anti-TPN antibody [EPR25083-20] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPN antibody [EPR25083-20] - BSA and Azide free (AB288574)

This data was developed using ab288565, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human lung tissue labelling TPN with ab288565 at 1/500 (1.002 ug/ml) followed by ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Mainly cytoplasmic staining on the leukocytes of human lung. The section was incubated with ab288565 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : 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-TPN antibody [EPR25083-20] - BSA and Azide free (AB288574)

This data was developed using ab288565, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon carcinom tissue labelling TPN with ab288565 at 1/500 (1.002 ug/ml) followed by ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Cytoplasmic staining on the cancer cells and stromal cells of human colon carcinoma (PMID : 26310568). The section was incubated with ab288565 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : 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-TPN antibody [EPR25083-20] - BSA and Azide free (AB288574)

This data was developed using ab288565, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Jurkat (Human T cell leukemia T lymphocyte) cells labelling TPN with ab288565 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 Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPN antibody [EPR25083-20] - BSA and Azide free (AB288574)

This data was developed using ab288565, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human lung adenocarcinoma tissue labelling TPN with ab288565 at 1/500 (1.002 ug/ml) followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection) was used. Cytoplasmic staining on the cancer cells and stromal cells of human lung adenocarcinoma. The section was incubated with ab288565 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : 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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TPN antibody [EPR25083-20] - BSA and Azide free (AB288574)

This data was developed using ab288565, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat cells labelling TPN with ab288565 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Jurkat cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 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 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TPN antibody [EPR25083-20] - BSA and Azide free (AB288574)

This data was developed using ab288565, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling TPN with ab288565 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 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 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPN antibody [EPR25083-20] - BSA and Azide free (AB288574)

This data was developed using ab288565, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling TPN with ab288565 at 1/500 (1.002 ug/ml) followed by ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Cytoplasmic staining on the stromal cells of human colon (PMID : 26310568). The section was incubated with ab288565 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : 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.

• WB

Supplier Data

Western blot - Anti-TPN antibody [EPR25083-20] - BSA and Azide free (AB288574)

This data was developed using ab288565, the same antibody clone in a different buffer formulation.

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

Upregulating the expression of TPN protein by IFN gamma treatment (PMID : 11169257)

All lanes:

Western blot - Anti-TPN antibody [EPR25083-20] (<a href='/en-us/products/primary-antibodies/tpn-antibody-epr25083-20-ab288565'>ab288565</a>) at 1/1000 dilution

Lane 1:

Untreated HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate at 20 µg

Lane 2:

HeLa treated with 100 ng/ml IFN gamma for 16 hours, 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: 48 kDa

Observed band size: 48 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-TPN antibody [EPR25083-20] - BSA and Azide free (AB288574)

This data was developed using ab288565, the same antibody clone in a different buffer formulation.

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

Exposure times :

Lanes 1 & 2 : 48 seconds

Lane 3 : 3 minutes

Lane 4 : 26 seconds

Laned 5 & 6 : 15 seconds

All lanes:

Western blot - Anti-TPN antibody [EPR25083-20] (<a href='/en-us/products/primary-antibodies/tpn-antibody-epr25083-20-ab288565'>ab288565</a>) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

Lane 4:

HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate at 20 µg

Lane 5:

A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 6:

Jurkat (human T cell leukemia T lymphocyte), 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: 48 kDa

Observed band size: 48 kDa

false

• WB

Supplier Data

Western blot - Anti-TPN antibody [EPR25083-20] - BSA and Azide free (AB288574)

This data was developed using ab288565, the same antibody clone in a different buffer formulation.

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

Upregulating the expression of TPN protein by IFN gamma treatment (PMID : 11169257)

All lanes:

Western blot - Anti-TPN antibody [EPR25083-20] (<a href='/en-us/products/primary-antibodies/tpn-antibody-epr25083-20-ab288565'>ab288565</a>) at 1/1000 dilution

Lane 1:

Untreated B16-F10 (Mouse skin melanoma), whole cell lysate at 10 µg

Lane 2:

B16-F10 treated with 100 ng/ml IFN gamma for 16 hours, whole cell lysate at 10 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: 48 kDa

Observed band size: 48 kDa

false

Exposure time: 48s