703605

antibody from Invitrogen Antibodies

Targeting: UBN1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

703605

Provider

Invitrogen Antibodies

Product name

Ubinuclein 1 Recombinant Rabbit Monoclonal Antibody (3H52L47)

Antibody type

Monoclonal

Antigen

Other

Description

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

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

3H52L47

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-Ubinuclein 1 Recombinant Rabbit Monoclonal Antibody (Product # 703605) and a 140kDa band corresponding to Ubinuclein 1 was observed across the cell lines tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), U-2 OS (Lane 2), MCF-7 (Lane 3), Hep G2 (Lane 4) and HT-29 (Lane 5) 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, HeLa cells were fixed and permeabilized with 0.5% Triton X-100 for detection of endogenous Ubinuclein 1 using Anti-Ubinuclein 1 Recombinant Rabbit Monoclonal Antibody (Product # 703605, 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 Ubinuclein1 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 nuclear localization of Ubinuclein 1. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin Immunoprecipitation (ChIP) assay of endogenous Ubinuclein 1 protein using Ubinuclein 1 Antibody: ChIP was performed using Anti-Ubinuclein 1 Recombinant Rabbit Monoclonal Antibody (Product # 703605, 5 µg) on sheared chromatin from 2 million HeLa cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to promoter regions of RBL1, CDK4 genes, gene bodies of GAPDH, ACTB and SATA satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.