PA1-32440

antibody from Invitrogen Antibodies

Targeting: PARK7 DJ-1, DJ1, GATD2

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Recommended by provider)

Antibody Data

Product number

PA1-32440

Provider

Invitrogen Antibodies

Product name

DJ-1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

PA1-32440 detects DJ1 from mouse, human, rat, hamster and fish samples.

Reactivity

Human, Mouse, Rat, Bovine, Chicken/Avian, Hamster, Zebrafish

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

Conc. Not Determined

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Identification of potential therapeutic compounds for Parkinson's disease using Drosophila and human cell models.

Sanz FJ, Solana-Manrique C, Muñoz-Soriano V, Calap-Quintana P, Moltó MD, Paricio N
Free radical biology & medicine 2017 Jul;108:683-691

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of HEK293 cells overexpressin DJ-1-V5 tag protein and mouse brain lysates using DJ-1 Polyclonal Antibody (Product # PA1-32440).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of DJ-1 was performed by loading human brain tissue lysate. Proteins were transferred to a membrane and probed with a DJ-1 Polyclonal Antibody (Product # PA1-32440).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of DJ-1 was achieved by transfecting HeLa cells with DJ-1 specific siRNAs (Silencer® select Product # s22304). Western blot analysis (Fig. a) was performed using whole cell extracts from the DJ-1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with DJ-1 Polyclonal Antibody (Product # PA1-32440, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25 µg/mL, 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 DJ-1.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using DJ-1 Polyclonal Antibody (Product # PA1-32440) and a 20 kDa band corresponding to DJ-1 was observed in U-2 OS, HeLa, Mouse Skeletal Muscle and Rat Skeletal Muscle and increased upon Hydrogen peroxide treatment in SH-SY5Y. Whole cell extracts (30 µg lysate) of SH-SY5Y (Lane 1), SH-SY5Y treated with Hydrogen peroxide (25 mM for 24 hours) (Lane 2), U-2 OS (Lane 3), HeLa (Lane 4) and tissue extracts (30 µg lysate) of Mouse Skeletal Muscle (Lane 5) and Rat Skeletal Muscle (Lane 6) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). 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).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of DJ1 in human cortex using a DJ1 polyclonal antibody (Product # PA1-32440).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of DJ-1 was performed using 70% confluent log phase He-La 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 1 hour at room temperature. The cells were labeled with DJ-1 Polyclonal Antibody (Product # PA1-32440) at 1:500 dilution 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 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry (Paraffin) analysis of DJ-1 in human cortex tissue using DJ-1 Polyclonal Antibody (Product # PA1-32440).