Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Loading Control

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Loading Control (AB7671)

ab7671 staining alpha 1 Sodium Potassium ATPase in Hek293 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab7671 at 1μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a single confocal section is shown.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Loading Control (AB7671)

IHC image of ab7671 staining in Human Kidney Carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab7671, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• IHC-P

AbReview46464****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Loading Control (AB7671)

ab7671 staining alpha 1 Sodium Potassium ATPase in human formaldehyde tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% BSA for 12 hours at 4°C; antigen retrieval was by heat mediation in a buffer pH 9. Samples were incubated with primary antibody (5μg/ml in 5% BSA) for 16 hours at 4°C. An Alexa Fluor^® 488-conjugated donkey anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

This image is courtesy of an Abreview submitted by Anne Sailer

• ICC

AbReview79962****

Immunocytochemistry - Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Loading Control (AB7671)

Immunocytochemistry analysis of formaldehyde-fixed mouse hepatocytes permeabilized with 0.2% Triton X-100 in PBS staining with ab7671 at 1/50 dilution. Secondary antibody was Alexa Fluor™ 488 Donkey anti-Ms. Samples were incubated with the primary antibody with 1% donkey serum in PBST for 3 hours at 22°C. Blocking was done using 1% donkey serum in PBST for 1 hour at 22°C.

This image is courtesy of an Abreview submitted by Armen Petrosyan

• WB

Project

Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Loading Control (AB7671)

Lanes 1 - 3:

Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Loading Control (ab7671) at 1 µg/mL

Lanes 4 - 6:

Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Loading Control (ab7671) at 2 µg/mL

Lanes 7 - 9:

Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Loading Control (ab7671) at 5 µg/mL

Lanes 1, 4 and 7:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 20 µg

Lanes 2, 5 and 8:

Brain (Mouse) Tissue Lysate (ab27253) at 20 µg

Lanes 3, 6 and 9:

Brain (Rat) Tissue Lysate (<a href='/en-us/products/unavailable/brain-rat-tissue-lysate-ab7942'>ab7942</a>) at 20 µg

Predicted band size: 113 kDa

true

Exposure time: 3min

• WB

Lab

Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Loading Control (AB7671)

Western blot analysis detecting Na, K-ATP-ase (alpha) in porcine proximal tubule protein, using ab7671. Band is at 112 kDa.

All lanes:

Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Loading Control (ab7671) at 1/5000 dilution

All lanes:

Porcine proximal tubule lysate

Predicted band size: 113 kDa

false

• WB

Project

Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Loading Control (AB7671)

All lanes:

alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Marker at 10 µg/mL

Lane 1:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg

Lane 2:

Human kidney tissue lysate - total protein (<a href='/en-us/products/unavailable/human-kidney-tissue-lysate-total-protein-ab30203'>ab30203</a>) at 20 µg

Lane 3:

Heart (Rabbit) Whole Cell Lysate - normal tissue (<a href='/en-us/products/unavailable/heart-rabbit-whole-cell-lysate-normal-tissue-ab29072'>ab29072</a>) at 20 µg

Lane 4:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 20 µg

Lane 5:

Brain (Human) Membrane Lysate - adult normal tissue at 20 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 113 kDa

Observed band size: 100 kDa

true

Exposure time: 3min