PA5-56878

antibody from Invitrogen Antibodies

Targeting: ATP6V1B1 ATP6B1, RTA1B, VATB, Vma2, VPP3

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-56878

Provider

Invitrogen Antibodies

Product name

ATP6V1B1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Immunogen sequence: MAMEIDSRPG GLPGSSCNLG AAREHMQAVT RNYITHPRVT YRTVCSVNGP LVVLDRVKFA QYAEIV

Concentration

0.1 mg/mL

References

Submitted references

A new Elf5(Cre)(ERT)(2-)(GFP) BAC transgenic mouse model for tracing Elf5 cell lineages in adult tissues.

Singh S, Elenio E, Leu NA, Romano RA, Vaughan AE, DeRiso J, Surendran K, Chakrabarti R
FEBS letters 2019 May;593(10):1030-1039

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of V-ATPase B1 in Lane 1: Marker (kDa) 230, 130, 95, 72, 56, 36, 28, 17, 11; Lane 2: Human cell line RT-4; Lane 3: Human cell line U-251MG sp. Samples were probed using a V-ATPase B1 Polyclonal Antibody (Product # PA5-56878).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-V-ATPase B1 Polyclonal Antibody (Product # PA5-56878) and a 56 kDa band corresponding to V-ATPase B1 was observed across cell lines tested. Membrane enriched extracts (30 µg lysate) of SK-BR-3 (Lane 1), HaCaT (Lane 2), HT-29 (Lane 3), BeWo (Lane 4), IMR-32 (Lane 5), Raji (Lane 6) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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 (1:1500) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005). A non-specific band picked up below 40 kDa marker has been marked with an asterisk.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of V-ATPase B1 was performed using 70% confluent log phase HaCaT and BeWo cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with V-ATPase B1 Polyclonal Antibody (Product # PA5-56878) at 1:100 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), (1:2500), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing endosomal localization. Panel e represents BeWo cells where the expression of the target protein is relatively low. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of V-ATPase B1 in human kidney and liver tissues using V-ATPase B1 Polyclonal Antibody (Product # PA5-56878). Corresponding ATP6V1B1 RNA-seq data are presented for the same tissues.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of V-ATPase B1 in human kidney using V-ATPase B1 Polyclonal Antibody (Product # PA5-56878) shows moderate to strong cytoplasmic positivity in cells in distal tubules.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of V-ATPase B1 in human salivary gland using V-ATPase B1 Polyclonal Antibody (Product # PA5-56878) shows strong cytoplasmic positivity in ductal cells.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of V-ATPase B1 in human breast using V-ATPase B1 Polyclonal Antibody (Product # PA5-56878) shows moderate cytoplasmic positivity in glandular cells.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of V-ATPase B1 in human liver using V-ATPase B1 Polyclonal Antibody (Product # PA5-56878) shows no positivity in hepatocytes as expected.