MA5-17016

antibody from Invitrogen Antibodies

Targeting: KPNA2 IPOA1, QIP2, RCH1, SRP1alpha

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-17016

Provider

Invitrogen Antibodies

Product name

KPNA2 Monoclonal Antibody (2G7)

Antibody type

Monoclonal

Antigen

Other

Description

MA5-17016 reacts with karyopherin 2 in human samples.

Antibody clone number

2G7

Concentration

1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of KPNA2 was achieved by transfecting HEK-293 cells with KPNA2 specific siRNAs (Silencer® select Product # s7921, s7920). Western blot analysis (Fig. a) was performed using whole cell extracts from the KPNA2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed using KPNA2 Monoclonal Antibody (Product # MA5-17016) and by F(ab)2-Rabbit anti-Rat IgG (H+L) Secondary Antibody, HRP (Product # PA1-29927, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to KPNA2.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Importin subunit alpha-1 (KPNA2) Monoclonal Antibody (Product # MA5-17016) on whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), A549 (Lane 2), HeLa (Lane 3), NIH/3T3 (Lane 4), PC-12 (Lane 5), LNCaP (Lane 6), Mouse Testis (Lane 7), Mouse Thymus (Lane 8), Mouse colon (Lane 9), Mouse Spleen (Lane 10) and Rat Testis (Lane 11). A 57.86 kDa band corresponding to Importin subunit alpha-1 (KPNA2) was observed except in Mouse colon which is reported to be negative. 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 F(ab)2-Rabbit anti-Rat IgG (H+L) Secondary Antibody, HRP (Product # PA1-29927, 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

Immunofluorescence analysis of KPNA2 was performed using HEK-293 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 1 hour at room temperature. The cells were labeled with KPNA2 Monoclonal Antibody (Product # MA5-17016) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A-11006) at a dilution of 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 composite image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.