PA5-55719

antibody from Invitrogen Antibodies

Targeting: NUP153 HNUP153

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-55719

Provider

Invitrogen Antibodies

Product name

NUP153 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Immunogen sequence: GQNREQRESG FSYPNFSLPA ANGLSSGVGG GGGKMRRERT RFVASKPLEE EEMEVPVLPK ISLPITSSSL PTFNFSSPEI TTSSPSPINS SQALTNKVQM TSP Highest antigen sequence identity to the following orthologs: Mouse - 79%, Rat - 79%.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.1 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

Knockdown of NUP153 was achieved by transfecting HeLa with NUP153 specific siRNAs (Silencer® select Product # s19374). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the NUP153 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with NUP153 Polyclonal Antibody (Product # PA5-55719, 0.1 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knockdown confirms that antibody is specific to NUP153.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-NUP153 Polyclonal Antibody (Product # PA5-55719) and a 200 kDa band corresponding to NUP153 was observed across cell lines tested. Nuclear enriched extracts (30 µg lysate) of HeLa (Lane 1) and K-562 (Lane 2) 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 (0.1 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). 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

Immunofluorescent staining of NUP153 in human cell line U-251 MG shows positivity in nuclear membrane. Samples were probed using a NUP153 Polyclonal Antibody (Product # PA5-55719).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of NUP153 was performed using 70% confluent log phase HeLa 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 NUP153 Polyclonal Antibody (Product # PA5-55719) at 2 µg/mL 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), (1:2000), 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 nuclear membrane and nucleus localization. Panel e 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

Immunohistochemistry analysis in human testis and pancreas tissues using HPA027896 antibody. Corresponding NUP153 RNA-seq data are presented for the same tissues. Samples were incubated in NUP153 polyclonal antibody (Product # PA5-55719).