Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] - Plasma Membrane Loading Control

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] - Plasma Membrane Loading Control (AB2872)

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human colon carcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1 : 200 with a mouse monoclonal antibody recognizing Sodium/Potassium ATPase alpha-1 ab2872 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] - Plasma Membrane Loading Control (AB2872)

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human kidney tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1 : 100 with a mouse monoclonal antibody recognizing Sodium/Potassium ATPase alpha-1 ab2872 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] - Plasma Membrane Loading Control (AB2872)

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1 : 50 with a mouse monoclonal antibody recognizing Sodium/Potassium ATPase alpha-1 ab2872 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

• Flow Cyt

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Flow Cytometry - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] - Plasma Membrane Loading Control (AB2872)

Overlay histogram showing HEK293 cells stained with ab2872 (red line). The cells were fixed with 100% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2872, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 4% paraformaldehyde used under the same conditions.

Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] - Plasma Membrane Loading Control (AB2872)

Immunocytochemistry/Immunofluorescence analysis of alpha 1 Sodium Potassium ATPase (green) in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 15 minutes at room temperature and blocked with 0.3% BSA for 15 minutes at room temperature. Cells were incubated with ab2872 (1 : 100) for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (1 : 500) for 30 minutes at room temperature. F-actin (red) was stained with DyLight 594 Phalloidin and nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.

• WB

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Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] - Plasma Membrane Loading Control (AB2872)

The 100 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to alpha 1 Sodium Potassium ATPase.

All lanes:

Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] - Plasma Membrane Loading Control (ab2872) at 1/500 dilution

Lane 1:

Human brain normal tissue lysate - membrane extract (<a href='/en-us/products/unavailable/human-brain-normal-tissue-lysate-membrane-extract-ab29456'>ab29456</a>) at 10 µg

Lane 2:

Human testis tissue lysate - total protein (<a href='/en-us/products/unavailable/human-testis-tissue-lysate-total-protein-ab30257'>ab30257</a>) at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/5000 dilution

Predicted band size: 113 kDa

Observed band size: 100 kDa,25 kDa,70 kDa

true

Exposure time: 8min

• WB

Supplier Data

Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] - Plasma Membrane Loading Control (AB2872)

Samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel. Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System.

Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit.

All lanes:

Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] - Plasma Membrane Loading Control (ab2872) at 1/1000 dilution

Lane 1:

Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 30 µg

Lane 2:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 30 µg

Lane 3:

MDA-MB-231 (Human breast adenocarcinoma cell line) whole cell lysate at 30 µg

Secondary

All lanes:

Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP at 1/4000 dilution

Predicted band size: 113 kDa

false

• IHC-P

AbReview10357****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] - Plasma Membrane Loading Control (AB2872)

ab2872 staining pig hepatocyte tissue sections by IHC-P. The section was fixed with Bouins and subjected to heat mediated antigen retrieval (at pH 9) prior to incubating with the primary antibody, diluted 1/2000, for 1 hour at 20°C. A Cy3® conjugated goat anti-mouse IgG antibody was used as the secondary.

This image is courtesy of an anonymous Abreview