- PA5-11274 - Invitrogen Antibodies IFITM3 antibody

Submitted references Increased Expression of Interferon-Induced Transmembrane 3 (IFITM3) in Stroke and Other Inflammatory Conditions in the Brain.

Harmon E, Doan A, Bautista-Garrido J, Jung JE, Marrelli SP, Kim GS
International journal of molecular sciences 2022 Aug 10;23(16)

Cutting Edge: NOX2 NADPH Oxidase Controls Infection by an Intracellular Bacterial Pathogen through Limiting the Type 1 IFN Response.

Rojas Márquez JD, Li T, McCluggage ARR, Tan JMJ, Muise A, Higgins DE, Brumell JH
Journal of immunology (Baltimore, Md. : 1950) 2021 Jan 15;206(2):323-328

Constitutively Expressed IFITM3 Protein in Human Endothelial Cells Poses an Early Infection Block to Human Influenza Viruses.

Sun X, Zeng H, Kumar A, Belser JA, Maines TR, Tumpey TM
Journal of virology 2016 Dec 15;90(24):11157-11167

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis using an IFITM3 polyclonal antibody (Product # PA5-11274) in HeLa, 293, A549, NCI-H460, HepG2, Sk-Br-3, A375 cell lysates (35 µg per lane).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis using an IFITM3 polyclonal antibody (Product # PA5-11274) in HeLa, 293, A549, NCI-H460, HepG2, Sk-Br-3, A375 cell lysates (35 µg per lane).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-IFITM3 Polyclonal Antibody (Product # PA5-11274) and a 14 kDa band corresponding to Interferon-induced transmembrane protein 3 was observed across cell lines.(30 µg lysate) of MCF7 (Lane 1), SH-SY5Y (Lane 2), SH-SY5Y (Lane 3), Caco-2 (Lane 4), Rat Lung (Lane 11) 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 (1:1000) 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Interferon-induced transmembrane protein 3 was achieved by transfecting MCF7 with Interferon-induced transmembrane protein 3 specific siRNAs (Silencer® select Product # S195033, S195034). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Interferon-induced transmembrane protein 3 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with IFITM3 Polyclonal Antibody (Product # PA5-11274, 1:1000 ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). 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 transmembrane protein 3.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of HeLa cells stained with an IFITM3 polyclonal antibody (Product # PA5-11274). HeLa cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min), then incubated with an IFITM3 polyclonal antibody (Product # PA5-11274) (1:200, 2 hr at room temperature). Primary antibody detected by fluor-conjugated donkey anti-rabbit secondary antibody (green) was used (1:1000, 1hr). Cytoplasmic actin was counterstained with red dye conjugated phalloidin (5.25 uM, 25 min). Nuclei were counterstained with Hoechst 33342 (blue) (10 µg/mL, 3 min)

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Interferon-induced transmembrane protein 3 was performed using 70% confluent log phase SH-SY5Y IFN gamma, 100 units/mL for 24 hrs. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with IFITM3 Polyclonal Antibody (Product # PA5-11274) 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 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 Cytoplasmic localization. Panel e represents no signal was observed Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: The IFITM3 protein level is increased by TNFalpha and IFNgamma in primary microglia. Primary microglia were treated with TNFalpha and IFNgamma (20 ng/mL) for 24 h. Western blotting was performed to measure protein level of IFITM3. ( A ) IFITM3 (seen at 14 kDa) was increased in the treatment group, compared to control. ( B ) Quantification of IFITM3 to beta-actin showed the significant induction of IFITM3 protein in microglia ( n = 6, ** p < 0.01, two-tailed unpaired Student's t -test).

PA5-11274

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