700229

antibody from Invitrogen Antibodies

Targeting: IRF7

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

700229

Provider

Invitrogen Antibodies

Product name

IRF7 Recombinant Rabbit Monoclonal Antibody (1H31L43)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

1H31L43

Vial size

100 µg

Concentration

0.5 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 analysis was performed on whole cell extracts (20 µg lysate) of Jurkat (Lane 1), HT-29 (Lane 2) and Raji (Lane 3). The blots were probed with Anti-IRF-7 Recombinant Rabbit Monoclonal Antibody (Product # 700229, 0.5-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 55 kDa band corresponding to IRF-7 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of IRF-7 in Jurkat cell lysate using an IRF-7 recombinant rabbit monoclonal antibody (Product # 700229) at a dilution of 1 µg/mL. NBT/BCIP was used as the substrate (Product # WB7105).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

CRISPR-Cas9 mediated genome editing ofIRF7 (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR847242_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of IRF7 was performed by loading 30 µg of A549 Cas9 (Lane 1) and A549 Cas9 cells transduced with IRF7 Lentiviral sgRNA (Lane 2) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 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 IRF7 Recombinant Rabbit Monoclonal Antibody (1H31L43) (Product # 700229, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:5000 dilution).Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). A loss of signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific toIRF7 (Fig (b)).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-IRF7 Recombinant Rabbit Monoclonal Antibody (1H31L43) (Product # 700229) and a 55kDa band corresponding to Interferon regulatory factor 7 was observed across cell lines. Whole cell extracts (30 µg lysate) of MCF7 (Lane 1), Raji (Lane 2), HEK-293 (Lane 3), HEK-293 (IFN-a treated (500 units/mL for 12 hrs )(Lane 4), HeLa (Lane 5), HeLa (IFN-a treated (500 units/mL for 12 hrs) (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).