702653

antibody from Invitrogen Antibodies

Targeting: RPS6KA5 MSK1, RLPK

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

702653

Provider

Invitrogen Antibodies

Product name

MSK1 Recombinant Rabbit Monoclonal Antibody (4H15L51)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Monkey, Pig, Rat, and Mouse.

Antibody clone number

4H15L51

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Nuclear extracts (30 µg lysate) of MCF-7 (Lane 1) and MDA-MB-231 (Lane 2). The blots were probed with Anti-MSK1 Recombinant Rabbit Monoclonal Antibody (Product # 702653, 1 µg/mL). A 120 kDa band corresponding to MSK1 was observed across the cell lines tested. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, Dexamethasone treated (1µM 2h) A549 cells were fixed and permeabilized for detection of endogenous MSK1 using Anti-MSK1 Recombinant Rabbit Monoclonal Antibody (Product # 702653, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of MSK1 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). 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 MSK1. Panel e) shows cytoplasmic translocation of MSK1 upon dexamethasone treatment. Panel f) represents control cells with no primary antibody to assess background.