- PA5-27627 - Invitrogen Antibodies CFL1 antibody

Submitted references Proteomic analysis of filaggrin deficiency identifies molecular signatures characteristic of atopic eczema.

Elias MS, Long HA, Newman CF, Wilson PA, West A, McGill PJ, Wu KC, Donaldson MJ, Reynolds NJ
The Journal of allergy and clinical immunology 2017 Nov;140(5):1299-1309

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Cofilin 1 (non-muscle) using A) 30 µg PC-12 whole cell lysate and B) 30 µg Rat-2 whole cell lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a Cofilin 1 (non-muscle) polyclonal antibody (Product # PA5-27627) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Cofilin 1 (non-muscle) using 30 µg of A431 lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a Cofilin 1 (non-muscle) polyclonal antibody (Product # PA5-27627) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Cofilin 1 (non-muscle) using 30 µg of NIH-3T3 lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a Cofilin 1 (non-muscle) polyclonal antibody (Product # PA5-27627) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of Cofilin was performed by separating 30 µg of various whole cell extracts by 12% SDS PAGE. Proteins were transferred to a membrane and probed with a Cofilin Polyclonal Antibody (Product # PA5-27627) at a dilution of 1:10000. A: Jurkat, B: Raji.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Cofilin Polyclonal Antibody (Product # PA5-27627). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with Cofilin Polyclonal Antibody (Product # PA5-27627) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Cofilin Polyclonal Antibody (Product # PA5-27627). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with Cofilin Polyclonal Antibody (Product # PA5-27627) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Cofilin Polyclonal Antibody (Product # PA5-27627). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with Cofilin Polyclonal Antibody (Product # PA5-27627) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Cofilin Polyclonal Antibody (Product # PA5-27627). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with Cofilin Polyclonal Antibody (Product # PA5-27627) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Cofilin was achieved by transfecting HeLa cells with Cofilin specific siRNAs (Silencer® select Product # s2037, s2936 ). Western blot analysis (Fig. a) was performed using Whole cell extracts from the untransfected cells (lane 1), non-targeting scrambled siRNA transfected cells (lane 2) and Cofilin knockdown cells (lane 3). The blot was probed with Cofilin Polyclonal Antibody (Product # PA5-27627, 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 Cofilin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-Cofilin Polyclonal Antibody(Product # PA5-27627) and a 18kDa band corresponding to Cofilin was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A-431 (Lane 2), Hep G2 (Lane 3), PC-12 (Lane 4) and NIH/3T3 (Lane 5) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). 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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Cofilin 1 (non-muscle) in paraformaldehyde-fixed HeLa cells using a Cofilin 1 (non-muscle) polyclonal antibody (Product # PA5-27627) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (Red) at a 1:2000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of Cofilin Cofilin in HeLa cells mock (left) and treated with 0.012 N NaOH/PBS for 1 min (right). Cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: Cofilin Polyclonal Antibody (Product # PA5-27627) diluted at 1:1000. Red: alpha Tubulin, a cytoskeleton marker, stained by alpha Tubulin antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of Cofilin 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 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Cofilin Polyclonal Antibody (Product # PA5-27627) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034), (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: Blue) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoskeleton, plasma membrane, nucleus and cytoplasm localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60 magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Cofilin Polyclonal Antibody detects CFL1 protein at cytosol on HBL435 xenograft by immunohistochemical analysis. Sample: Paraffin-embedded HBL435 xenograft. Cofilin Polyclonal Antibody (Product # PA5-27627) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

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

Supportive validation

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

PA5-27627

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