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Parkin Recombinant Antibody [PSH08-11] - Guinea pig IgG2 (Chimeric)

Usd: 385

Specification

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Catalog# HA601663

Parkin Recombinant Antibody [PSH08-11] - Guinea pig IgG2 (Chimeric)

Application

  • WB

  • IF-Cell

  • IHC-P

  • IF-Tissue

  • FC(Intra)

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

Parkin Recombinant Antibody [PSH08-11] - Guinea pig IgG2 (Chimeric)

Antibody Type

Recombinant Chimeric Antibody

Immunogen

Recombinant protein within human Parkin aa 1-200.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, IF-Tissue, FC(Intra)

Target Molecular Weight

Predicted band size: 52 kDa

Positive Control

SH-SY5Y (Human neuroblastoma cell) cell lysate, COS-1 (African green monkey kidney fibroblast) cell lysate, PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, Pig brain tissue lysate.

Conjugation

unconjugated

Clone Number

PSH08-11

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:2,000

  • IF-Tissue

  • 1:500

  • FC(Intra)

  • 1:1,000

Target

Function

Parkin is a 465-amino acid residue E3 ubiquitin ligase, a protein that in humans and mice is encoded by the PARK2 gene. Parkin plays a critical role in ubiquitination – the process whereby molecules are covalently labelled with ubiquitin (Ub) and directed towards degradation in proteasomes or lysosomes. Ubiquitination involves the sequential action of three enzymes. First, an E1 ubiquitin-activating enzyme binds to inactive Ub in eukaryotic cells via a thioester bond and mobilises it in an ATP-dependent process. Ub is then transferred to an E2 ubiquitin-conjugating enzyme before being conjugated to the target protein via an E3 ubiquitin ligase. There exists a multitude of E3 ligases, which differ in structure and substrate specificity to allow selective targeting of proteins to intracellular degradation.

Background References

1. Quinn PMJ et al. PINK1/PARKIN signalling in neurodegeneration and neuroinflammation. Acta Neuropathol Commun. 2020 Nov

2. Li J et al. PINK1/Parkin-mediated mitophagy in neurodegenerative diseases. Ageing Res Rev. 2023 Feb

Subcellular Location

Mitochondrion, mitochondrion outer membrane, endoplasmic reticulum, cytosol, Nucleus, neuron projection, postsynaptic density, presynapse.

UNIPROT

SwissProt: O60260 Human

SwissProt: Q9WVS6 Mouse

SwissProt: Q9JK66 Rat

Synonyms

AR JP antibody

E3 ubiquitin ligase antibody

E3 ubiquitin protein ligase parkin antibody

E3 ubiquitin-protein ligase parkin antibody

FRA6E antibody

LPRS 2 antibody

LPRS2 antibody

PARK 2 antibody

Park2 antibody

Parkin 2 antibody

Expand

Images

  • Western blot analysis of Parkin on different lysates with Guinea pig anti-Parkin antibody (<a href="/products/HA601663" style="font-weight: bold;text-decoration: underline;">HA601663</a>) at 1/5,000 dilution. <br /> <br />Lane 1: SH-SY5Y (Human neuroblastoma cell) cell lysate <br />Lane 2: COS-1 (African green monkey kidney fibroblast) cell lysate <br />Lane 3: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate <br />Lane 4: Mouse brain tissue lysate <br />Lane 5: Rat brain tissue lysate <br />Lane 6: Pig brain tissue lysate <br /> <br />Lysates/proteins at 10 µg/Lane. <br />Exposure time: 10 seconds; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA601663" style="font-weight: bold;text-decoration: underline;">HA601663</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Guinea pig IgG-HRP (<a href="/products/HA1021" style="font-weight: bold;text-decoration: underline;">HA1021</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 52 kDa <br />Observed band size: 52 kDa

    Western blot analysis of Parkin on different lysates with Guinea pig anti-Parkin antibody (HA601663) at 1/5,000 dilution.

    Lane 1: SH-SY5Y (Human neuroblastoma cell) cell lysate
    Lane 2: COS-1 (African green monkey kidney fibroblast) cell lysate
    Lane 3: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate
    Lane 4: Mouse brain tissue lysate
    Lane 5: Rat brain tissue lysate
    Lane 6: Pig brain tissue lysate

    Lysates/proteins at 10 µg/Lane.
    Exposure time: 10 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA601663, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Guinea pig IgG-HRP (HA1021), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 52 kDa
    Observed band size: 52 kDa

  • Application: Immunocytochemistry (IF-cell) <br /> <br />Species: Mouse <br />Sample: Neuro-2a (Mouse brain neuroblastoma cell) <br /> <br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. <br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature. <br />Antibody dilution buffer: 1% BSA in PBST. <br /> <br />Primary antibody: <a href="/products/HA601663" style="font-weight: bold;text-decoration: underline;">HA601663</a>, 1/100, overnight at 4°C. <br />Secondary antibody: Goat Anti-Guinea pig IgG (iFluor&trade; 594, <a href="/products/HA1143" style="font-weight: bold;text-decoration: underline;">HA1143</a>), 45 hours at room temperature. <br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, Green), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

    Application: Immunocytochemistry (IF-cell)

    Species: Mouse
    Sample: Neuro-2a (Mouse brain neuroblastoma cell)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.

    Primary antibody: HA601663, 1/100, overnight at 4°C.
    Secondary antibody: Goat Anti-Guinea pig IgG (iFluor™ 594, HA1143), 45 hours at room temperature.
    Counterstain: Beta tubulin (HA601187, Green), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Mouse <br />Tissue: Cerebral cortex <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA601663" style="font-weight: bold;text-decoration: underline;">HA601663</a>, 1/2,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1021" style="font-weight: bold;text-decoration: underline;">HA1021</a>, 30 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Cerebral cortex
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA601663, 1/2,000, 1 hour at room temperature.
    Secondary antibody: HA1021, 30 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Rat <br />Tissue: Cerebral cortex <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA601663" style="font-weight: bold;text-decoration: underline;">HA601663</a>, 1/2,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1021" style="font-weight: bold;text-decoration: underline;">HA1021</a>, 30 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Rat
    Tissue: Cerebral cortex
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA601663, 1/2,000, 1 hour at room temperature.
    Secondary antibody: HA1021, 30 minutes at room temperature.

  • Application: Immunofluorescence (IF-tissue) <br /> <br />Species: Mouse <br />Tissue: Cerebral cortex <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature. <br />Primary antibody: <a href="/products/HA601663" style="font-weight: bold;text-decoration: underline;">HA601663</a>, 1/500, overnight at 4°C. <br />Secondary antibody: Goat Anti-Guinea pig IgG (iFluor&trade; 594, <a href="/products/HA1143" style="font-weight: bold;text-decoration: underline;">HA1143</a>), 1.5 hours at room temperature.

    Application: Immunofluorescence (IF-tissue)

    Species: Mouse
    Tissue: Cerebral cortex
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
    Primary antibody: HA601663, 1/500, overnight at 4°C.
    Secondary antibody: Goat Anti-Guinea pig IgG (iFluor™ 594, HA1143), 1.5 hours at room temperature.

  • Application: Flow Cytometry (Intra) <br /> <br />Species: Mouse <br />Sample: Neuro-2a (Mouse brain neuroblastoma cell) <br /> <br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. <br />Permeabilization: 0.1% Tween-20, 15 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature. <br />Antibody dilution buffer: 1x PBS. <br /> <br />Primary antibody: <a href="/products/HA601663" style="font-weight: bold;text-decoration: underline;">HA601663</a> (1/1,000, Red), 15 minutes at room temperature. <br />Secondary antibody: Goat Anti-Guinea pig IgG (iFluor&trade; 594, <a href="/products/HA1143" style="font-weight: bold;text-decoration: underline;">HA1143</a>), 15 hours at room temperature.

    Application: Flow Cytometry (Intra)

    Species: Mouse
    Sample: Neuro-2a (Mouse brain neuroblastoma cell)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
    Antibody dilution buffer: 1x PBS.

    Primary antibody: HA601663 (1/1,000, Red), 15 minutes at room temperature.
    Secondary antibody: Goat Anti-Guinea pig IgG (iFluor™ 594, HA1143), 15 hours at room temperature.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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