MA5-15040

antibody from Invitrogen Antibodies

Targeting: RPS6KA3 CLS, HU-3, MRX19, RSK2

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Antibody Data

Product number

MA5-15040

Provider

Invitrogen Antibodies

Product name

RSK1/RSK2/RSK3 Monoclonal Antibody (F.940.7)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody.

Reactivity

Human, Mouse, Rat, Guinea Pig

Host

Rabbit

Isotype

IgG

Antibody clone number

F.940.7

Vial size

100 µL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of Ribosomal protein S6 kinase alpha-1 was achieved by transfecting HEK-293 with Ribosomal protein S6 kinase alpha-1 specific siRNAs (Silencer® select Product # S12274, S12273). Western Blot analysis (Fig. a) was performed using Whole cell extracts from the Ribosomal protein S6 kinase alpha-1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The Blot was probed with RSK1/RSK2/RSK3 Monoclonal Antibody (F.940.7) (Product # MA5-15040, 1:1000) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). Densitometric analysis of this western Blot is shown in histogram (Fig. b).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot was performed using Anti-RSK1/RSK2/RSK3 Monoclonal Antibody (F.940.7) (Product # MA5-15040) and a 80 kDa band corresponding to Ribosomal protein S6 kinase alpha-1 was observed. Whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), HeLa (Lane 2), K-562 (Lane 3), A-431 (Lane 4), HEL 92.1.7 (Lane 5) and BJ (Lane 6) 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 (1:1000) 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 Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of RSK1/RSK2/RSK3 in HeLa cells using a RSK1/RSK2/RSK3 monoclonal antibody (Product # MA5-15040) (green). Actin filaments are labeled with a fluorescent red phalloidin.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of RSK1/RSK2/RSK3 was performed using 70% confluent log phase U-87 MG 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 RSK1/RSK2/RSK3 Monoclonal Antibody (F.940.7) (Product # MA5-15040) 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), (1:2,000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominant cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7C1115LZR).