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Phospho-PPP1CA (T320) Recombinant Rabbit Monoclonal Antibody [PSH10-27] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751344

Phospho-PPP1CA (T320) Recombinant Rabbit Monoclonal Antibody [PSH10-27] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

Phospho-PPP1CA (T320) Recombinant Rabbit Monoclonal Antibody [PSH10-27] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Thr320 of Human PPP1CA.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 38 kDa

Positive Control

HeLa treated with 100nM Calyculin A for 30 minutes cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate, human breast cancer tissue, human testis tissue, rat testis tissue.

Conjugation

unconjugated

Clone Number

PSH10-27

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:20,000

  • IP

  • 1-2μg/sample

Target

Function

Serine/threonine-protein phosphatase PP1-alpha catalytic subunit is an enzyme that in humans is encoded by the PPP1CA gene. The protein encoded by this gene is one of the three catalytic subunits of protein phosphatase 1 (PP1). PP1 is a serine/threonine specific protein phosphatase known to be involved in the regulation of a variety of cellular processes, such as cell division, glycogen metabolism, muscle contractility, protein synthesis, and HIV-1 viral transcription. Increased PP1 activity has been observed in the end stage of heart failure. Studies in both human and mice suggest that PP1 is an important regulator of cardiac function. Three alternatively spliced transcript variants encoding different isoforms have been found for this gene.

Background References

1. Zhang L et al. Arginine methylation of PPP1CA by CARM1 regulates glucose metabolism and affects osteogenic differentiation and osteoclastic differentiation. Clin Transl Med. 2023 Sep

2. Liang H et al. Lysine methylation of PPP1CA by the methyltransferase SUV39H2 disrupts TFEB-dependent autophagy and promotes intervertebral disc degeneration. Cell Death Differ. 2023 Sep

Subcellular Location

Cytoplasm, Nucleus, nucleoplasm, nucleolus.

UNIPROT

SwissProt: P62136 Human

SwissProt: P62137 Mouse

SwissProt: P62138 Rat

Synonyms

Alpha isoform serine threonine protein phosphatase PP1alpha 1 catalytic subunit antibody

Catalytic subunit antibody

EC 3.1.3.16 antibody

MGC15877 antibody

MGC1674 antibody

PP 1A antibody

PP-1A antibody

PP1A antibody

PP1A_HUMAN antibody

PP1alpha antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-PPP1CA (T320) on different lysates with Rabbit anti-Phospho-PPP1CA (T320) antibody (<a href="/products/HA751344" style="font-weight: bold;text-decoration: underline;">HA751344</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate<br />Lane 3: NIH/3T3 cell lysate<br />Lane 4: PC-12 cell lysate<br />Lane 5: PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate<br />Lane 6: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 38 kDa<br />Observed band size: 35 kDa<br /><br />Exposure time: 3 minutes; 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/HA751344" style="font-weight: bold;text-decoration: underline;">HA751344</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.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-PPP1CA (T320) on different lysates with Rabbit anti-Phospho-PPP1CA (T320) antibody (HA751344) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate
    Lane 3: NIH/3T3 cell lysate
    Lane 4: PC-12 cell lysate
    Lane 5: PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate
    Lane 6: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 38 kDa
    Observed band size: 35 kDa

    Exposure time: 3 minutes; 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 (HA751344) 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.

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Phospho-PPP1CA (T320) antibody (<a href="/products/HA751344" style="font-weight: bold;text-decoration: underline;">HA751344</a>) at 1/20,000 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/HA751344" style="font-weight: bold;text-decoration: underline;">HA751344</a>) at 1/20,000 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 breast cancer tissue with Rabbit anti-Phospho-PPP1CA (T320) antibody (HA751344) at 1/20,000 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 (HA751344) at 1/20,000 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 testis tissue with Rabbit anti-Phospho-PPP1CA (T320) antibody (<a href="/products/HA751344" style="font-weight: bold;text-decoration: underline;">HA751344</a>) at 1/20,000 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/HA751344" style="font-weight: bold;text-decoration: underline;">HA751344</a>) at 1/20,000 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 testis tissue with Rabbit anti-Phospho-PPP1CA (T320) antibody (HA751344) at 1/20,000 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 (HA751344) at 1/20,000 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 testis tissue with Rabbit anti-Phospho-PPP1CA (T320) antibody (<a href="/products/HA751344" style="font-weight: bold;text-decoration: underline;">HA751344</a>) at 1/20,000 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/HA751344" style="font-weight: bold;text-decoration: underline;">HA751344</a>) at 1/20,000 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 testis tissue with Rabbit anti-Phospho-PPP1CA (T320) antibody (HA751344) at 1/20,000 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 (HA751344) at 1/20,000 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Phospho-PPP1CA (T320) was immunoprecipitated from 0.2 mg HeLa treated with 50mM Calyculin A for 3 hours cell lysate with <a href="/products/HA751344" style="font-weight: bold;text-decoration: underline;">HA751344</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA751344" style="font-weight: bold;text-decoration: underline;">HA751344</a> at 1/2,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary 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: HeLa treated with 50mM Calyculin A for 3 hours cell lysate (input)<br />Lane 2: <a href="/products/HA751344" style="font-weight: bold;text-decoration: underline;">HA751344</a> IP in HeLa treated with 50mM Calyculin A for 3 hours cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA751344" style="font-weight: bold;text-decoration: underline;">HA751344</a> in HeLa treated with 50mM Calyculin A for 3 hours cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 1 minute 23 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    ☑ Cell treatment (CT)

    Phospho-PPP1CA (T320) was immunoprecipitated from 0.2 mg HeLa treated with 50mM Calyculin A for 3 hours cell lysate with HA751344 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751344 at 1/2,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa treated with 50mM Calyculin A for 3 hours cell lysate (input)
    Lane 2: HA751344 IP in HeLa treated with 50mM Calyculin A for 3 hours cell lysate
    Lane 3: Rabbit IgG instead of HA751344 in HeLa treated with 50mM Calyculin A for 3 hours cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 1 minute 23 seconds; ECL: K1801

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

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