MA5-16397

antibody from Invitrogen Antibodies

Targeting: RPS6 S6

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Recommended by provider)

Other assay (Recommended by provider)

Antibody Data

Product number

MA5-16397

Provider

Invitrogen Antibodies

Product name

Phospho-S6 (Ser235, Ser236) Monoclonal Antibody (SP50)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 30 min at room temperature. A suggested positive control is breast ductal carcinoma.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

SP50

Vial size

500 µL

Storage

4° C, do not freeze

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Phospho-S6 (Ser235, Ser236) Monoclonal Antibody (SP50)(Product # MA5-16397) and a 32 kDa band corresponding to Phospho-S6 (Ser235, Ser236) was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of U-2 OS (Lane 1), U-2 OS treated with IGF (Lane 2), U-2 OS treated with Rapamycin (Lane 3), U-2 OS treated with both IGF and Rapamycin (Lane 4), HEK-293 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) 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). IGF was used at 50 ng/mL concentration for 6 hrs and Rapamycin was used at 50 nM strength for 4 hrs.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of Phospho-S6 Ribosomal pSer235/236 using anti-Phospho-S6 Ribosomal pSer235/236 Monoclonal Antibody (Product # MA5-16397) in Breast Carcinoma Cancer Tissue. The recommened dilution for this antibody in immunohistochemistry applications is 1:100.