PA5-105027

antibody from Invitrogen Antibodies

Targeting: MAP3K5 ASK1, MAPKKK5, MEKK5

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-105027

Provider

Invitrogen Antibodies

Product name

Phospho-ASK1 (Ser966) Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Antibody detects endogenous levels of ASK1 only when phosphorylated at Serine 966.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

1 mg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Phospho-ASK1 (Ser966) Polyclonal Antibody (Product # PA5-105027) and a 170 kDa band corresponding to Mitogen-activated protein kinase kinase kinase 5 was observed across tested panel. Whole cell extracts (40 µg lysate) of MCF7 (Lane 1), MCF7 treated with Nocodazole 200 ng/mL for 16 hours (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Phospho-ASK1 (Ser966) in rat brain/mouse brain/ (absence (-) or presence (+) of non-phospho and phospho peptide). Samples were incubated with Phospho-ASK1 (Ser966) polyclonal antibody (Product # PA5-105027).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Phospho-ASK1 (Ser966) in MDA-MB-435. Samples were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100, blocked with 10% serum (45 min at 25°C) incubated with Phospho-ASK1 (Ser966) polyclonal antibody (Product # PA5-105027) using a dilution of 1:200 (1 hr, 37°C), and followed by goat anti-rabbit IgG Alexa Fluor 594 at a dilution of 1:600.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of paraffin-embedded Phospho-ASK1 (Ser966) in human Lymph node tissue sections. Antigen retrieval was performed using citrate buffer. Samples were blocked with blocking buffer (1.5 hr, 22°C), incubated with Phospho-ASK1 (Ser966) polyclonal antibody (Product # PA5-105027) using a dilution of 1:100 (1.5 hr, 22°C), followed by HRP conjugated goat anti-rabbit.