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PARK7/DJ1 Recombinant Rabbit Monoclonal Antibody [SN07-21]

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Specification

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Catalog# ET1611-45

PARK7/DJ1 Recombinant Rabbit Monoclonal Antibody [SN07-21]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Predicted reactivity

  • Cynomolgus monkey

  • Pig

Predicted species support after-sales service
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

PARK7/DJ1 Recombinant Rabbit Monoclonal Antibody [SN07-21]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human PARK7 aa 11-60 / 189.

Species Reactivity

Human, Mouse, Rat (Predicted: Cynomolgus monkey, Pig)

Predicted species support after-sales service

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, IP, FC

Molecular Weight

Predicted band size: 20 kDa

Positive Control

HeLa cell lysate, SH-SY5Y cell lysate, HepG2 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, A549 cell lysate, HeLa, NIH/3T3, C6, human brain tissue, mouse cerebrum tissue, mouse hippocampus tissue, rat brain tissue.

Conjugation

unconjugated

Clone Number

SN07-21

RRID

AB_3070024

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000-1:5,000

  • IF-Cell

  • 1:200

  • IF-Tissue

  • 1:500

  • IHC-P

  • 1:5,000

  • FC

  • 1:50-1:100

  • IP

  • Use at an assay dependent concentration.

Target

Function

PARK7/DJ1 is a ubiquitously expressed protein involved in various cellular processes including cell proliferation, RNA-binding, and oxidative stress. The protein has been found to colocalize within a subset of pathologic tau inclusions in a diverse group of neurodegenerative disorders known as tauopathies. Defects in PARK7/DJ1 are the cause of autosomal recessive early-onset Parkinson's disease 7 (PARK7). Parkinson's disease (PD) is a complex, multifactorial disorder that typically manifests after the age of 50 years. The disease is characterized by bradykinesia, resting tremor, muscular rigidity and postural instability. The pathology involves the loss of dopaminergic neurons in the substantia nigra and the presence of Lewy bodies (intraneuronal accumulations of aggregated proteins), in surviving neurons in various areas of the brain. PARK7 is characterized by onset before 40 years and slow progression. It has also been suggested that PARK7/DJ1 is a mitogen dependent oncogene product involved in Ras related signal transduction pathways.

Background References

1. Marín-Buera L et al. Differential proteomic profiling unveils new molecular mechanisms associated with mitochondrial complex III deficiency. J Proteomics 113:38-56 (2015).

2. Bifsha P et al. Rgs6 is required for adult maintenance of dopaminergic neurons in the ventral substantia nigra. PLoS Genet 10:e1004863 (2014).

Sequence Similarity

Belongs to the peptidase C56 family.

Tissue Specificity

Highly expressed in pancreas, kidney, skeletal muscle, liver, testis and heart. Detected at slightly lower levels in placenta and brain (at protein level). Detected in astrocytes, Sertoli cells, spermatogonia, spermatids and spermatozoa. Expressed by pancreatic islets at higher levels than surrounding exocrine tissues.

Post-translational Modification

Sumoylated on Lys-130 by PIAS2 or PIAS4; which is enhanced after ultraviolet irradiation and essential for cell-growth promoting activity and transforming activity.; Cys-106 is easily oxidized to sulfinic acid.; Undergoes cleavage of a C-terminal peptide and subsequent activation of protease activity in response to oxidative stress.

Subcellular Location

Cell membrane, Cytoplasm, Endoplasmic reticulum, Membrane, Mitochondrion, Nucleus.

UNIPROT

SwissProt: Q99497 Human

SwissProt: Q99LX0 Mouse

SwissProt: O88767 Rat

Synonyms

CAP1 antibody

DJ-1 antibody

DJ1 antibody

DJ1 protein antibody

Epididymis secretory sperm binding protein Li 67p antibody

FLJ27376 antibody

FLJ34360 antibody

FLJ92274 antibody

HEL S 67p antibody

Oncogene DJ1 antibody

Expand

Images

  • Western blot analysis of PARK7/DJ1 on different lysates with Rabbit anti-PARK7/DJ1 antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: SH-SY5Y cell lysate<br />Lane 3: HepG2 cell lysate<br />Lane 4: Mouse brain tissue lysate<br />Lane 5: Rat brain tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 20 kDa<br />Observed band size: 20 kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of PARK7/DJ1 on different lysates with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: SH-SY5Y cell lysate
    Lane 3: HepG2 cell lysate
    Lane 4: Mouse brain tissue lysate
    Lane 5: Rat brain tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 20 kDa
    Observed band size: 20 kDa

    Exposure time: 20 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-45) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of PARK7/DJ1 on different lysates with Rabbit anti-PARK7/DJ1 antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549-si NT cell lysate<br />Lane 2: A549-si PARK7/DJ1 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 20 kDa<br />Observed band size: 20 kDa<br /><br />Exposure time: 30 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of PARK7/DJ1 on different lysates with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/2,000 dilution.

    Lane 1: A549-si NT cell lysate
    Lane 2: A549-si PARK7/DJ1 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 20 kDa
    Observed band size: 20 kDa

    Exposure time: 30 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-45) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling PARK7/DJ1 with Rabbit anti-PARK7/DJ1 antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/200 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PARK7/DJ1 antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling PARK7/DJ1 with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/200 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling PARK7/DJ1 with Rabbit anti-PARK7/DJ1 antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/200 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PARK7/DJ1 antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of NIH/3T3 cells labeling PARK7/DJ1 with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/200 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of C6 cells labeling PARK7/DJ1 with Rabbit anti-PARK7/DJ1 antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/200 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PARK7/DJ1 antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of C6 cells labeling PARK7/DJ1 with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/200 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-PARK7/DJ1 antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-45) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue with Rabbit anti-PARK7/DJ1 antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-45) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue with Rabbit anti-PARK7/DJ1 antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-45) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-PARK7/DJ1 antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1611-45" style="font-weight: bold;text-decoration: underline;">ET1611-45</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-45) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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Citation