PA5-31254

antibody from Invitrogen Antibodies

Targeting: IFIT1 G10P1, GARG-16, IFI56, IFNAI1

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-31254

Provider

Invitrogen Antibodies

Product name

IFIT1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: HeLa, HeLa(10 ng/mL hIFN-gamma1 treatment for 16hr).

Concentration

1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of IFIT1 using 30 µg of HCT116 lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with an IFIT1 polyclonal antibody (Product # PA5-31254) at a dilution of 1:1000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of IFIT1 was performed by separating 30 µg of untreated (–) and treated (+) HeLa whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a IFIT1 Polyclonal Antibody (Product # PA5-31254) at a dilution of 1:1000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of Interferon-induced protein with tetratricopeptide repeats 1 was achieved by transfecting HeLa treated with Human IFN Beta (1000 U/mL for 24hrs) with Interferon-induced protein with tetratricopeptide repeats 1 specific siRNAs (Silencer® select Product # s7150, s7151). For detectable levels of IFIT1, transfected cells were treated with Human IFN Beta. Western blot analysis (Fig. a) was performed using Whole cell extracts from the Interferon-induced protein with tetratricopeptide repeats 1 knockdown cells treated with Human IFN Beta (lane 6), IFIT3 control HeLa cells (Lane 5), non-targeting scrambled siRNA transfected HeLa cells treated with Human IFN Beta (lane 4), non-targeting scrambled siRNA transfected HeLa cells (lane 3), Untransfected HeLa cells treated with Human IFN Beta (Lane 2) and untransfected HeLa cells (lane 1).The blot was probed with IFIT1 Polyclonal Antibody (Product # PA5-31254, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Interferon-induced protein with tetratricopeptide repeats 1.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-IFIT1 Polyclonal Antibody (Product # PA5-31254) and a 55 kDa band corresponding to Interferon-induced protein with tetratricopeptide repeats 1 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HT-1080 (Lane 1), HT-1080 treated with Human IFN Beta (1000U/mL for 24 hrs) (Lane 2), HeLa (Lane 3) and HeLa treated with Human IFN Beta (1000U/mL for 24 hrs) (Lane 4) 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, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).