PA5-17344

antibody from Invitrogen Antibodies

Targeting: ACIN1 ACINUS, fSAP152, KIAA0670

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-17344

Provider

Invitrogen Antibodies

Product name

Acinus Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

35.1 µg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Acinus in extracts from Jurkat and A204 cell lines using Acinus polyclonal antibody (Product # PA5-17344).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot was performed using Anti-Acinus Polyclonal Antibody (Product # PA5-17344) and ~220 kDa, 150 kDa, 75-80 kDa bands corresponding to ACIN1 was observed across cell lines tested and generated p45 and p36 alternate splice forms upon apoptotic stimulus. Whole cell extracts (40 µg lysate) of Hep G2 (Lane 1), untreated Jurkat (Lane 2), staurosporine- treated Jurkat (Lane 3), camptothecin-treated Jurakt (Lane 4), untreated U-2 OS (Lane 5), and etoposide-treated U-2 OS (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 (1:1250 dilution) and detected by using chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10000 dilution) using 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

Immunofluorescence analysis of ACIN1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Acinus Polyclonal Antibody (Product # PA5-17344) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:2500 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization for ACIN1. Panel e represents control HeLa cells with no primary antibody to assess background. The images were captured at 60X magnification