Anti-NCAM1 antibody [EPR26939-108] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR26939-108] - BSA and Azide free (AB313780)

This data was developed using ab313779, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling NCAM1 with ab313779 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human liver.The section was incubated with ab313779 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed 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-NCAM1 antibody [EPR26939-108] - BSA and Azide free (AB313780)

This data was developed using ab313779, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human appendix tissue labeling NCAM1 with ab313779 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on stroma of human appendix (PMID : 27015206).The section was incubated with ab313779 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed 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-NCAM1 antibody [EPR26939-108] - BSA and Azide free (AB313780)

This data was developed using ab313779, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling NCAM1 with ab313779 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human cerebrum (PMID : 19246644).The section was incubated with ab313779 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed 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-NCAM1 antibody [EPR26939-108] - BSA and Azide free (AB313780)

This data was developed using ab313779, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human ovarian carcin tissue labeling NCAM1 with ab313779 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human ovarian carcinoma.The section was incubated with ab313779 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed 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-NCAM1 antibody [EPR26939-108] - BSA and Azide free (AB313780)

This data was developed using ab313779, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human astrocytoma tissue labeling NCAM1 with ab313779 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human astrocytoma.The section was incubated with ab313779 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NCAM1 antibody [EPR26939-108] - BSA and Azide free (AB313780)

This data was developed using ab313779, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized Human PBMC (human primary peripheral blood mononuclear cell) cells labelling NCAM1 with ab313779 at 1/500 (1.016 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 membranous staining in subsets of human PBMCs, which is not co-localized with CD3.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). Anti-human CD3 mouse monoclonal antibody (Alexa Fluor® 647) was used to counterstain tubulin at 1/200 2.5 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

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Immunocytochemistry/ Immunofluorescence - Anti-NCAM1 antibody [EPR26939-108] - BSA and Azide free (AB313780)

This data was developed using ab313779, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) cells labelling NCAM1 with ab313779 at 1/500 (1.016 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 membranous staining in No-GFP-CD16.NK-92 cell line.Negative control : MCF7 (PMID : 30050775).Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/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.

• Flow Cyt

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Flow Cytometry - Anti-NCAM1 antibody [EPR26939-108] - BSA and Azide free (AB313780)

This data was developed using ab313779, the same antibody clone in a different buffer formulation. Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling NCAM1 with ab313779 at 1/500 dilution (0.1 ug)/Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Cells are co-stained with CD3 conjugated to Alexa Fluor®647. Gated on viable cells.

• Flow Cyt

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Flow Cytometry - Anti-NCAM1 antibody [EPR26939-108] - BSA and Azide free (AB313780)

This data was developed using ab313779, the same antibody clone in a different buffer formulation. Flow cytometric analysis of MCF7 (human breast adenocarcinoma epithelial cell, Left) - / No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) (Right) cells labelling NCAM1 with ab313779 at 1/500 dilution (0.1 ug)/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). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Negative control : MCF7 (PMID : 30050775). Gated on viable cells.

• WB

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Western blot - Anti-NCAM1 antibody [EPR26939-108] - BSA and Azide free (AB313780)

This data was developed using ab313779, the same antibody clone in a different buffer formulation.

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

Negative control : MCF7 (PMID : 18772882).

The 120, 140 and 180kDa bands are different isoforms as reported in the literature (PMID : 26288071)

The identity of the lower MW bands at approximately 75 kDa and 50kDa are unknown.

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

Exposure time : Lane 1 : 147 seconds; Lane 2, 3 : 92 seconds.

All lanes:

Western blot - Anti-NCAM1 antibody [EPR26939-108] (<a href='/en-us/products/primary-antibodies/ncam1-antibody-epr26939-108-ab313779'>ab313779</a>) at 1/1000 dilution

Lane 1:

No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92&reg;) cell with no GFP tag) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

SH-SY5Y (human neuroblastoma epithelial cell) 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

Observed band size: 120 kDa,140 kDa,180 kDa

false

• WB

Supplier Data

Western blot - Anti-NCAM1 antibody [EPR26939-108] - BSA and Azide free (AB313780)

This data was developed using ab313779, the same antibody clone in a different buffer formulation.

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

The 120, 140 and 180kDa bands are different isoforms as reported in the literature (PMID : 26288071).

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-NCAM1 antibody [EPR26939-108] (<a href='/en-us/products/primary-antibodies/ncam1-antibody-epr26939-108-ab313779'>ab313779</a>) at 1/1000 dilution

Lane 1:

Human hippocampus tissue lysate at 20 µg

Lane 2:

Human cerebellum tissue 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

Observed band size: 120 kDa,140 kDa,180 kDa

false

Exposure time: 15s