- PA5-23072 - Invitrogen Antibodies PINK1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of PINK1 in 1.0 mg/mL HeLa lysate. Samples were incubated in PINK1 polyclonal antibody (Product # PA5-23072) using a dilution of 1:50. This experiment was performed under reducing conditions using the 12-230 k Da separation system. * Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of PINK1 was performed by loading 30 µg of U-2 OS (Lane 1) and U-2 OS - PINK1 knockout (Lane 2) Whole cell extracts. The blot was probed with Anti-PINK1 Polyclonal Antibody (Product # PA5-23072,1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to PINK1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-PINK1 Polyclonal Antibody (Product # PA5-23072) and a 64 kDa band corresponding to PINK1 was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of U-2 OS(Lane 1), HeLa (Lane 2), SH-SY5Y (Lane 3), U-87 MG (Lane 4) and PC-12 (Lane 5), were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) 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)
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Experimental details: Immunofluorescent analysis of PINK1 using a polyclonal antibody (Product # PA5-23072).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry analysis of PINK1 in HepG2 cells. Samples were incubated in PINK1 polyclonal antibody (Product # PA5-23072) followed by DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of PINK1 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 PINK1 Polyclonal Antibody (Product # PA5-23072) at 1:200 dilution 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 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 dilution). Panel d represents the merged image showing Mitochondrial localization. Panel e represents control HeLa cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of PINK1 in paraffin embedded mouse brain. Samples were incubated in PINK1 polyclonal antibody (Product # PA5-23072).

PA5-23072