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Phospho-Nucleolin (T76) Recombinant Rabbit Monoclonal Antibody [JE64-03]

Usd: 385

Specification

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Catalog# HA722942

Phospho-Nucleolin (T76) Recombinant Rabbit Monoclonal Antibody [JE64-03]

Application

  • WB

  • IHC-P

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

Phospho-Nucleolin (T76) Recombinant Rabbit Monoclonal Antibody [JE64-03]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Thr76 of Human Nucleolin.

Species Reactivity

Human

Validated Applications

WB, IHC-P

Target Molecular Weight

Predicted band size: 77 kDa

Positive Control

K-562 cell lysate, K-562 treated with 50ng/mL Calyculin A for 20 minutes cell lysate, human tonsil tissue, human kidney cancer tissue.

Conjugation

unconjugated

Clone Number

JE64-03

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*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

  • IHC-P

  • 1:200

Target

Function

Nucleolin is the major nucleolar protein of growing eukaryotic cells. It is found associated with intranucleolar chromatin and pre-ribosomal particles. It induces chromatin decondensation by binding to histone H1. It is thought to play a role in pre-rRNA transcription and ribosome assembly. May play a role in the process of transcriptional elongation. Binds RNA oligonucleotides with 5'-UUAGGG-3' repeats more tightly than the telomeric single-stranded DNA 5'-TTAGGG-3' repeats.

Background References

1. Ezzatifar F et al. Nucleolin; A tumor associated antigen as a potential lung cancer biomarker. Pathol Res Pract. 2022 Dec.

2. Cangelosi D et al. Nucleolin expression has prognostic value in neuroblastoma patients. EBioMedicine. 2022 Nov.

Subcellular Location

Nucleus, nucleolus;Cytoplasm

UNIPROT

SwissProt: P19338 Human

Synonyms

C23 antibody

FLJ45706 antibody

MS1116 antibody

NCL antibody

Nucl antibody

NUCL_HUMAN antibody

Nucleolin antibody

Protein C23 antibody

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-Nucleolin (T76) on different lysates with Rabbit anti-Phospho-Nucleolin (T76) antibody (<a href="/products/HA722942" style="font-weight: bold;text-decoration: underline;">HA722942</a>) at 1/2,000 dilution.<br /><br />Lane 1: K-562 cell lysate (20 µg/Lane)<br />Lane 2: K-562 treated with 50ng/mL Calyculin A for 20 minutes cell lysate (20 µg/Lane)<br />Lane 3: K-562 treated with 50ng/mL Calyculin A for 20 minutes cell lysate, the membrane treated with λpp for 1 hour (20 µg/Lane)<br />Predicted band size: 77 kDa<br />Observed band size: 105 kDa<br />Exposure time: 17 seconds; ECL: K1801;<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/HA722942" style="font-weight: bold;text-decoration: underline;">HA722942</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-Nucleolin (T76) on different lysates with Rabbit anti-Phospho-Nucleolin (T76) antibody (HA722942) at 1/2,000 dilution.

    Lane 1: K-562 cell lysate (20 µg/Lane)
    Lane 2: K-562 treated with 50ng/mL Calyculin A for 20 minutes cell lysate (20 µg/Lane)
    Lane 3: K-562 treated with 50ng/mL Calyculin A for 20 minutes cell lysate, the membrane treated with λpp for 1 hour (20 µg/Lane)
    Predicted band size: 77 kDa
    Observed band size: 105 kDa
    Exposure time: 17 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 (HA722942) 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;">☑ Cell treatment (CT)</span><br /><br />Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Phospho-Nucleolin (T76) antibody (<a href="/products/HA722942" style="font-weight: bold;text-decoration: underline;">HA722942</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA722942" style="font-weight: bold;text-decoration: underline;">HA722942</a>) at 1/200 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.

    ☑ Cell treatment (CT)

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Phospho-Nucleolin (T76) antibody (HA722942) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA722942) at 1/200 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 kidney cancer tissue with Rabbit anti-Phospho-Nucleolin (T76) antibody (<a href="/products/HA722942" style="font-weight: bold;text-decoration: underline;">HA722942</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA722942" style="font-weight: bold;text-decoration: underline;">HA722942</a>) at 1/200 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 kidney cancer tissue with Rabbit anti-Phospho-Nucleolin (T76) antibody (HA722942) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA722942) at 1/200 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.

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