PA5-31862

antibody from Invitrogen Antibodies

Targeting: S100A6 2A9, CABP, CACY, PRA

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-31862

Provider

Invitrogen Antibodies

Product name

S100A6 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Recommended positive controls: U87-MG. Predicted reactivity: Mouse (96%), Rat (95%), Pig (87%), Rabbit (90%), Chicken (88%), Rhesus Monkey (100%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.86 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using S100A6 Polyclonal Antibody (Product # PA5-31862). Sample (30 µg of whole cell lysate). Lane A: U87-MG . 15% SDS PAGE. S100A6 Polyclonal Antibody (Product # PA5-31862) diluted at 1:5,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-S100A6 Polyclonal Antibody (Product # PA5-31862) and a 10kDa band corresponding to Protein S100-A6 was observed. Whole cell extracts (30 µg lysate) of A549 (Lane 1), PC-3 (Lane 2), U-87 MG (Lane 3), HEK-293 (Lane 4), K-562 (Lane 5) and LNCaP (Lane 6) were electrophoresed using Novex™ 16% Tricine Protein Gel (Product # EC6695BOX). 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 (1:5000) 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). Band of interest was not observed in HEK-293, K-562 and LNCaP which are reported to be either negative or express S100A6 at very low levels.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of S100A6 in paraformaldehyde-fixed A431 cells using a S100A6 polyclonal antibody (Product # PA5-31862) at a 1:500 dilution.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of S100A6 was performed using 70% confluent log phase PC-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.01% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with S100A6 Polyclonal Antibody (Product # PA5-31862) 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), (1:2000), 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).