703383

antibody from Invitrogen Antibodies

Targeting: P2RY6 P2Y6

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

703383

Provider

Invitrogen Antibodies

Product name

P2Y6 Recombinant Rabbit Monoclonal Antibody (8H45L52)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Mouse, Rat, Bovine.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

8H45L52

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-P2Y6 Recombinant Rabbit Monoclonal Antibody (Product # 703383) and a 36 kDa band corresponding to P2Y6 was observed across cell lines tested. Membrane enriched extracts (50 µg lysate) of MCF-7 (Lane 1) and Hep G2 (Lane 2) were electrophoresed using Novex® NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 (1:1000 dilution) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 1:5000 dilution) using the iBright FL 1500 (Product # A44115). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-P2Y6 Recombinant Rabbit Monoclonal Antibody (Product # 703383) and a 36 kDa band corresponding to P2Y6 was observed in relevant cell line tested. Membrane enriched extract (30 µg lysate) of MCF-7 (Lane 1) and T47D (Lane2) were electrophoresed using Novex® NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 (1:1000 dilution) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 1:5000 dilution) ) using the iBright FL 1500 (Product # A44115). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, MCF-7 cells were fixed for the detection of endogenous P2Y6 using Anti-P2Y6 Recombinant Rabbit Monoclonal Antibody (Product # 703383, 1:100 dilution) and labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 conjugate (Product # A32731, 1:2000). Panel a) shows representative cells that were stained for detection and localization of P2Y6 protein (green), Panel b) is stained for nuclei (blue) using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating Membrane and cytoplasmic localization of P2Y6. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.