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PIWIL2 Recombinant Rabbit Monoclonal Antibody [PSH27-16]

Usd: 385

Specification

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Catalog# HA724677

PIWIL2 Recombinant Rabbit Monoclonal Antibody [PSH27-16]

Application

  • WB

  • IHC-Fr

  • IHC-P

  • IP

  • mIHC

Reactivity

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

PIWIL2 Recombinant Rabbit Monoclonal Antibody [PSH27-16]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within mouse PIWIL2 aa 1-250.

Species Reactivity

Mouse, Rat

Validated Applications

WB, IHC-Fr, IHC-P, IP, mIHC

Target Molecular Weight

Predicted band size: 109 kDa

Positive Control

Rat testis tissue lysate, Mouse testis tissue lysate.

Conjugation

unconjugated

Clone Number

PSH27-16

Reactivity Data

Tested Tested Tested
Verified (internally validated)
Published Tested
Reported in literature (not internally validated)
Predicted Tested
Predicted reactive (based on sequence homology)
Not recommended Tested
Not recommended (failed internal validation)
WB IHC-P IHC-Fr FC(Intra) IF-Cell IP mIHC
human
Not Recommended
 
 
Not Recommended
Not Recommended
 
 
mouse
Tested Tested
Tested Tested
Tested Tested
 
 
Tested Tested
Tested Tested
rat
Tested Tested
Tested Tested
 
 
 
 
 

Product Features

Form

Liquid

Concentration

0.5 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.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-Fr

  • 1:250

  • IHC-P

  • 1:500

  • IP

  • 1-2μg/sample

  • mIHC

  • 1:1,000

Target

Function

Endoribonuclease that plays a central role during spermatogenesis by repressing transposable elements and preventing their mobilization, which is essential for the germline integrity (By similarity). Plays an essential role in meiotic differentiation of spermatocytes, germ cell differentiation and in self-renewal of spermatogonial stem cells (By similarity). Acts via the piRNA metabolic process, which mediates the repression of transposable elements during meiosis by forming complexes composed of piRNAs and Piwi proteins and govern the methylation and subsequent repression of transposons (By similarity). During piRNA biosynthesis, plays a key role in the piRNA amplification loop, also named ping-pong amplification cycle, by acting as a 'slicer-competent' piRNA endoribonuclease that cleaves primary piRNAs, which are then loaded onto 'slicer-incompetent' PIWIL4 (By similarity). PIWIL2 slicing produces a pre-miRNA intermediate, which is then processed in mature piRNAs, and as well as a 16 nucleotide by-product that is degraded (By similarity). Required for PIWIL4/MIWI2 nuclear localization and association with secondary piRNAs antisense (By similarity). Besides their function in transposable elements repression, piRNAs are probably involved in other processes during meiosis such as translation regulation (By similarity). Indirectly modulates expression of genes such as PDGFRB, SLC2A1, ITGA6, GJA7, THY1, CD9 and STRA8 (By similarity). When overexpressed, acts as an oncogene by inhibition of apoptosis and promotion of proliferation in tumors. Represses circadian rhythms by promoting the stability and activity of core clock components BMAL1 and CLOCK by inhibiting GSK3B-mediated phosphorylation and ubiquitination-dependent degradation of these proteins.

Background References

1. Li Y et al. PIWIL2/PDK1 Axis Promotes the Progression of Cervical Epithelial Lesions via Metabolic Reprogramming to Maintain Tumor-Initiating Cell Stemness. Adv Sci (Weinh). 2024 Dec

2. Hammad G et al. HEPPAR1 and PIWIL2 as Panel Markers for Hepatocellular Carcinoma. Asian Pac J Cancer Prev. 2024 Jun

Subcellular Location

Cytoplasm.

UNIPROT

SwissProt: Q8CDG1 Mouse

Entrez Gene: 306011 Rat

Synonyms

Piwi-like protein 2

EC:3.1.26.

Piwil2

Mili

Images

  • mIHC analysis of mouse testis tissue (paraffin-embedded) with PIWIL2 (<a href="/products/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a>, 1/1,000) and Vimentin (<a href="/products/ET1610-39" style="font-weight: bold;text-decoration: underline;">ET1610-39</a>, 1/10,000).<br /><br />The immunostaining was performed with the IRISKitCmTSA Kit (<a href="/products/900802" style="font-weight: bold;text-decoration: underline;">900802</a>). DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

    mIHC analysis of mouse testis tissue (paraffin-embedded) with PIWIL2 (HA724677, 1/1,000) and Vimentin (ET1610-39, 1/10,000).

    The immunostaining was performed with the IRISKitCmTSA Kit (900802). DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

  • Application: Immunofluorescence (IHC-Fr)<br /><br />Species: Mouse<br />Tissue: Testis<br />Sample: Frozen section<br /><br />Antigen retrieval: Not required<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 /><br />Primary antibody: <a href="/products/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a>, 1/250, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 1.5 hours at room temperature.

    Application: Immunofluorescence (IHC-Fr)

    Species: Mouse
    Tissue: Testis
    Sample: Frozen section

    Antigen retrieval: Not required

    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: HA724677, 1/250, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Mouse <br />Tissue: Testis <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/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a>, 1/500, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Testis
    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: HA724677, 1/500, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Mouse <br />Tissue: Brain <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/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a>, 1/500, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature. <br /> <br />Negative expression of PIWIL2 protein in brain is consistent with the predicted expression pattern.

    ☑ Relative expression (RE)

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Brain
    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: HA724677, 1/500, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

    Negative expression of PIWIL2 protein in brain is consistent with the predicted expression pattern.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Rat <br />Tissue: Testis <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/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a>, 1/500, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Rat
    Tissue: Testis
    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: HA724677, 1/500, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Rat <br />Tissue: Brain <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/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a>, 1/500, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature. <br /> <br />Negative expression of PIWIL2 protein in brain is consistent with the predicted expression pattern.

    ☑ Relative expression (RE)

    Application: Immunohistochemistry (IHC-P)

    Species: Rat
    Tissue: Brain
    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: HA724677, 1/500, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

    Negative expression of PIWIL2 protein in brain is consistent with the predicted expression pattern.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of PIWIL2 on different lysates with Rabbit anti-PIWIL2 antibody (<a href="/products/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a>) at 1/2,000 dilution. <br /> <br />Lane 1: Rat testis tissue lysate <br />Lane 2: Rat brain tissue lysate <br /> <br />Negative control: Brain (PMID: 16377660). <br /> <br />Lysates/proteins at 30 µg/Lane. <br />Exposure time: 20 seconds; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a>, 1/2,000 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-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 109 kDa <br />Observed band size: 109 kDa

    ☑ Relative expression (RE)

    Western blot analysis of PIWIL2 on different lysates with Rabbit anti-PIWIL2 antibody (HA724677) at 1/2,000 dilution.

    Lane 1: Rat testis tissue lysate
    Lane 2: Rat brain tissue lysate

    Negative control: Brain (PMID: 16377660).

    Lysates/proteins at 30 µg/Lane.
    Exposure time: 20 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724677, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 109 kDa
    Observed band size: 109 kDa

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of PIWIL2 on different lysates with Rabbit anti-PIWIL2 antibody (<a href="/products/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a>) at 1/2,000 dilution. <br /> <br />Lane 1: Mouse testis tissue lysate <br />Lane 2: Mouse brain tissue lysate <br /> <br />Negative control: Brain (PMID: 16377660). <br /> <br />Lysates/proteins at 30 µg/Lane. <br />Exposure time: 20 seconds; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a>, 1/2,000 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-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 109 kDa <br />Observed band size: 109 kDa

    ☑ Relative expression (RE)

    Western blot analysis of PIWIL2 on different lysates with Rabbit anti-PIWIL2 antibody (HA724677) at 1/2,000 dilution.

    Lane 1: Mouse testis tissue lysate
    Lane 2: Mouse brain tissue lysate

    Negative control: Brain (PMID: 16377660).

    Lysates/proteins at 30 µg/Lane.
    Exposure time: 20 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724677, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 109 kDa
    Observed band size: 109 kDa

  • Immunoprecipitation (IP) <br /> <br />PIWIL2 was immunoprecipitated in 0.2 mg mouse testis tissue lysate with <a href="/products/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a> at 1/5,000 dilution. HRP Conjugated Rabbit IgG kappa light chain Mouse Monoclonal Antibody (<a href="/products/M1208-2" style="font-weight: bold;text-decoration: underline;">M1208-2</a>) at 1/5,000 dilution was used for 1 hour at room temperature. <br /> <br />Lane 1: Mouse testis tissue lysate (input) <br />Lane 2: <a href="/products/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a> IP in mouse testis tissue lysate <br />Lane 3: Rabbit IgG instead of <a href="/products/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a> in mouse testis tissue lysate <br /> <br />Exposure time: 1 minute 50 seconds <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Primary dilution: <a href="/products/HA724677" style="font-weight: bold;text-decoration: underline;">HA724677</a>, 1/2,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), 2 hours at room temperature <br /> <br />Predicted band size: 109 kDa <br />Observed band size: 109 kDa

    Immunoprecipitation (IP)

    PIWIL2 was immunoprecipitated in 0.2 mg mouse testis tissue lysate with HA724677 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724677 at 1/5,000 dilution. HRP Conjugated Rabbit IgG kappa light chain Mouse Monoclonal Antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: Mouse testis tissue lysate (input)
    Lane 2: HA724677 IP in mouse testis tissue lysate
    Lane 3: Rabbit IgG instead of HA724677 in mouse testis tissue lysate

    Exposure time: 1 minute 50 seconds
    Blocking: 5% NFDM/TBST, 1 hour at room temperature

    Primary dilution: HA724677, 1/2,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

    Predicted band size: 109 kDa
    Observed band size: 109 kDa

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