PA5-42697

antibody from Invitrogen Antibodies

Targeting: ATP6AP1 16A, Ac45, ATP6IP1, ATP6S1, CF2, ORF, VATPS1, XAP-3, XAP3

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-42697

Provider

Invitrogen Antibodies

Product name

ATP6AP1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Peptide sequence: SPVIHPPVSY NDTAPRILFW AQNFSVAYKD QWEDLTPLTF GVQELNLTGS

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of human OVCAR-3 cell lysate using an anti-ATP6AP1 polyclonal antibody (Product # PA5-42697).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-ATP6AP1 Polyclonal Antibody (Product # PA5-42697) and a 52-55kDa band corresponding to V-type proton ATPase subunit S1 was observed across all the cell lines and tissue samples tested. 42 kDa band was also observed in some cell lines which may correspond to a C-terminal cleaved fragment. Membrane enriched extracts (30 µg lysate) of SK-BR-3 (Lane 1), BeWo (Lane 2), MCF7 (Lane 3), THP-1 (Lane 4), NIH:OVCAR-3 (Lane 5), NTERA-2 cl.D1 (Lane 6), HEL 92.1.7 (Lane 7), NK-92 (Lane 8), Mouse Brain (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of V-type proton ATPase subunit S1 was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with ATP6AP1 Polyclonal Antibody (Product # PA5-42697) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:4000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing endoplasmic reticulum and golgi localizations. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7A1110LZR) and externally deconvoluted (D.Sage et al. / Methods 115 (2017) 28-41).