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Nestin Recombinant Rabbit Monoclonal Antibody [PSH07-89] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751196

Nestin Recombinant Rabbit Monoclonal Antibody [PSH07-89] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IHC-P

  • IHC-Fr

  • IF-Tissue

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

Nestin Recombinant Rabbit Monoclonal Antibody [PSH07-89] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within mouse Nestin aa 700-1,000.

Species Reactivity

Human, Mouse, Rat

Validated Applications

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

Molecular Weight

Predicted band size: 207 kDa

Positive Control

RD cell lysate, Neuro-2a cell lysate, Mouse brain (P0) tissue lysate, Mouse brain (P7) tissue lysate, Mouse brain tissue lysate, U-87 MG, C2C12, C6, human kidney tissue, mouse kidney tissue, rat kidney tissue, mouse embryonic brain tissue.

Conjugation

unconjugated

Clone Number

PSH07-89

Product Features

Form

Liquid

Concentration

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

  • IF-Cell

  • 1:1,000-1:5,000

  • IHC-P

  • 1:1,000

  • IHC-Fr

  • 1:500-1:2,000

  • IF-Tissue

  • 1:500

  • IP

  • Use at an assay dependent concentration.

Target

Function

Nestin is a major intermediate filament (IF) protein of embryonic central nervous system progenitor cells. It is also a component of the dynamic IF network during muscle development, where it polymerizes with Desmin and Vimentin. Nestin co-assembles with Vimentin or a-internexin and forms heterodimer coiled-coil molecules which then further assemble into 10 nml IFs. Deletion of the IF consensus rod domain in nestin alters nestin localization in CNS precursor cells and radial glial cells in vivo. Nestin is a marker for neuroepithelial stem cells, glioma cells and tumor endothelial cells during rapid growth. During axon elongation of differen-tiation neurons, nestin localizes to the growth cones and may play a role in growth cone guidance. In the rat adrenal gland, nestin is expressed by the zona fasciculata and the zona reticularis. Nestin is also expressed by dermatomal cells and by myoblasts during the earliest stages of myogenesis.

Background References

1. Chen H et al. Targeting Nestin(+) hepatic stellate cells ameliorates liver fibrosis by facilitating TbetaRI degradation. J Hepatol. 2021 May

2. Wang J et al. Nestin promotes pulmonary fibrosis via facilitating recycling of TGF-beta receptor I. Eur Respir J. 2022 May

Subcellular Location

Cytoplasm, Intermediate filament.

UNIPROT

SwissProt: P48681 Human

SwissProt: Q6P5H2 Mouse

SwissProt: P21263 Rat

Synonyms

ESTM 46 antibody

FLJ 21841 antibody

FLJ21841 antibody

Intermediate filament protein antibody

Nbla00170 antibody

nes antibody

NEST_HUMAN antibody

Nestin antibody

Images

  • Application: IHC-Fr<br /><br />Species: Mouse<br /><br />Site: E14.5 embryonic brain<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/500<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Mouse

    Site: E14.5 embryonic brain

    Sample: Frozen section

    Antibody concentration: 1/500

    Antigen retrieval: Not required

  • Application: IHC-Fr<br /><br />Species: Rat<br /><br />Site: Kindey<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/500<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Rat

    Site: Kindey

    Sample: Frozen section

    Antibody concentration: 1/500

    Antigen retrieval: Not required

  • Application: IF-tissue<br /><br />Species: Mouse<br /><br />Site: E14.5 embryonic brain<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    Application: IF-tissue

    Species: Mouse

    Site: E14.5 embryonic brain

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

  • Application: IF-tissue<br /><br />Species: Mouse<br /><br />Site: embryos (cartilage)<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    Application: IF-tissue

    Species: Mouse

    Site: embryos (cartilage)

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

  • Application: IF-tissue<br /><br />Species: Rat<br /><br />Site: brain<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    Application: IF-tissue

    Species: Rat

    Site: brain

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Nestin antibody (<a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/1,000 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/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/1,000 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 kidney tissue with Rabbit anti-Nestin antibody (HA751196) at 1/1,000 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 (HA751196) at 1/1,000 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 kidney tissue with Rabbit anti-Nestin antibody (<a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/1,000 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/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/1,000 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 kidney tissue with Rabbit anti-Nestin antibody (HA751196) at 1/1,000 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 (HA751196) at 1/1,000 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 kidney tissue with Rabbit anti-Nestin antibody (<a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/1,000 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/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/1,000 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 kidney tissue with Rabbit anti-Nestin antibody (HA751196) at 1/1,000 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 (HA751196) at 1/1,000 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 embryonic brain tissue with Rabbit anti-Nestin antibody (<a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/1,000 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/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/1,000 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 embryonic brain tissue with Rabbit anti-Nestin antibody (HA751196) at 1/1,000 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 (HA751196) at 1/1,000 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;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Nestin on different lysates with Rabbit anti-Nestin antibody (<a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/2,000 dilution.<br /><br />Lane 1: RD cell lysate<br />Lane 2: HEK-293 cell lysate (negative)<br />Lane 3: Neuro-2a cell lysate<br />Lane 4: Mouse brain (P0) tissue lysate<br />Lane 5: Mouse brain (P7) tissue lysate<br />Lane 6: Mouse brain tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 177/207 kDa<br />Observed band size: 150-300 kDa<br /><br />Exposure time: 15 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/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/2,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) 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.

    ☑ Relative expression (RE)

    Western blot analysis of Nestin on different lysates with Rabbit anti-Nestin antibody (HA751196) at 1/2,000 dilution.

    Lane 1: RD cell lysate
    Lane 2: HEK-293 cell lysate (negative)
    Lane 3: Neuro-2a cell lysate
    Lane 4: Mouse brain (P0) tissue lysate
    Lane 5: Mouse brain (P7) tissue lysate
    Lane 6: Mouse brain tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 177/207 kDa
    Observed band size: 150-300 kDa

    Exposure time: 15 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 (HA751196) at 1/2,000 dilution was used in primary antibody dilution (K1803) 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 Nestin on different lysates with Rabbit anti-Nestin antibody (<a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Nestin KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 177 kDa<br />Observed band size: 150-300 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/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/1,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) 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 Nestin on different lysates with Rabbit anti-Nestin antibody (HA751196) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Nestin KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 177 kDa
    Observed band size: 150-300 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 (HA751196) at 1/1,000 dilution was used in primary antibody dilution (K1803) 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 C2C12 cells labeling Nestin with Rabbit anti-Nestin antibody (<a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/5,000 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-Nestin antibody (<a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/5,000 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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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 C2C12 cells labeling Nestin with Rabbit anti-Nestin antibody (HA751196) at 1/5,000 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-Nestin antibody (HA751196) at 1/5,000 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 (HA601187, 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 Nestin with Rabbit anti-Nestin antibody (<a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/1,000 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-Nestin antibody (<a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a>) at 1/1,000 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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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 Nestin with Rabbit anti-Nestin antibody (HA751196) at 1/1,000 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-Nestin antibody (HA751196) at 1/1,000 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 (HA601187, 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.

  • Nestin was immunoprecipitated from 0.2 mg C2C12 cell lysate with <a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a> at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: C2C12 cell lysate (input)<br />Lane 2: <a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a> IP in C2C12 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA751196" style="font-weight: bold;text-decoration: underline;">HA751196</a> in C2C12 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 6 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    Nestin was immunoprecipitated from 0.2 mg C2C12 cell lysate with HA751196 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751196 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: C2C12 cell lysate (input)
    Lane 2: HA751196 IP in C2C12 cell lysate
    Lane 3: Rabbit IgG instead of HA751196 in C2C12 cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 6 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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