Anti-NCS1 antibody [EPR7699]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-NCS1 antibody [EPR7699] (AB129166)

Overlay histogram showing SH-SY5Y cells stained with ab129166 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab129166 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCS1 antibody [EPR7699] (AB129166)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labelling NCS1 with ab248324 at 1/10000 (0.101 μg/ml) dilution followed by a LeicaDS9800 (Bond^™ Polymer Refine Detection) at a ready to use dilution. Positive staining on human cerebrum. The section was incubated with ab248324 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Incubate 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 LeicaDS9800 (Bond^™ Polymer Refine Detection) at a ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. This data was developed using ab248324, the same antibody clone in a different buffer formulation.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCS1 antibody [EPR7699] (AB129166)

ab129165 at 1/250 dilution staining NCS1 in paraffin-embedded Human brain tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCS1 antibody [EPR7699] (AB129166)

Immunohistochemical analysis of paraffin-embedded human liver tissue labelling NCS1 with ab248324 at 1/10000 (0.101 μg/ml) dilution followed by a LeicaDS9800 (Bond^™ Polymer Refine Detection) at a ready to use dilution. Negative control : no staining on human liver. The section was incubated with ab248324 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with hematoxylin. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. This data was developed using ab248324, the same antibody clone in a different buffer formulation.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCS1 antibody [EPR7699] (AB129166)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labelling NCS1 with ab248324 at 1/10000 (0.101 μg/ml) dilution followed by a LeicaDS9800 (Bond^™ Polymer Refine Detection) at a ready to use dilution. Positive staining on mouse cerebrum. The section was incubated with ab248324 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Incubate 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 LeicaDS9800 (Bond^™ Polymer Refine Detection) at a ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. This data was developed using ab248324, the same antibody clone in a different buffer formulation.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCS1 antibody [EPR7699] (AB129166)

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labelling NCS1 with ab248324 at 1/10000 (0.101 μg/ml) dilution followed by a LeicaDS9800 (Bond^™ Polymer Refine Detection) at a ready to use dilution. Positive staining on rat cerebrum. The section was incubated with ab248324 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Incubate 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 LeicaDS9800 (Bond^™ Polymer Refine Detection) at a ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. This data was developed using ab248324, the same antibody clone in a different buffer formulation.

• WB

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Western blot - Anti-NCS1 antibody [EPR7699] (AB129166)

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

All lanes:

Western blot - Anti-NCS1 antibody [EPR7699] (ab129166) at 1/5000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Rat brain 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 1/20000 dilution

Predicted band size: 22 kDa

Observed band size: 21 kDa

false

Exposure time: 180s

• WB

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Western blot - Anti-NCS1 antibody [EPR7699] (AB129166)

All lanes:

Western blot - Anti-NCS1 antibody [EPR7699] (ab129166) at 1/1000 dilution

Lane 1:

U-87 MG cell lysates at 10 µg

Lane 2:

SH-SY5Y cell lysates at 10 µg

Lane 3:

A549 cell lysates at 10 µg

Lane 4:

293T cell lysates at 10 µg

Lane 5:

Fetal brain tissue lysates at 10 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/2000 dilution

Predicted band size: 22 kDa

false