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NDRG1 Recombinant Rabbit Monoclonal Antibody [JM32-02]

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Specification

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Catalog# ET1705-20

NDRG1 Recombinant Rabbit Monoclonal Antibody [JM32-02]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

  • IHC-Fr

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

NDRG1 Recombinant Rabbit Monoclonal Antibody [JM32-02]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human NDRG1 aa 345-394 / 394.

Species Reactivity

Human, Mouse, Rat

Validated Applications

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

Molecular Weight

Predicted band size: 43 kDa

Positive Control

PC-3 cell lysate, SW480 cell lysate, RPMI 8226 cell lysate, RAW264.7 cell lysate, Mouse eyeball tissue lysate, Rat eyeball tissue lysate, MCF-7, A431, human prostate tissue, mouse kidney tissue, human colon carcinoma tissue.

Conjugation

unconjugated

Clone Number

JM32-02

RRID

AB_3070553

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. 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

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:200-1:1,000

  • IP

  • 1:50

  • IHC-Fr

  • 1:200

Target

Function

Protein NDRG1 is a protein that in humans is encoded by the NDRG1 gene. This gene is a member of the N-myc downregulated gene family which belongs to the alpha/beta hydrolase superfamily. The protein encoded by this gene is a cytoplasmic protein involved in stress responses, hormone responses, cell growth, and differentiation[citation needed]. Mutations in this gene have been reported to be causative the autosomal-recessive version of Charcot-Marie-Tooth disease known as CMT4D. It has been reported that NDRG1 localizes to the endosomes and is a Rab4a effector involved in vesicular recycling. As reviewed by Fang et al., NDRG1 is involved in embryogenesis and development, cell growth and differentiation, lipid biosynthesis and myelination, stress responses, immunity, DNA repair and cell adhesion among other functions. NDRG1 is localised in the cytoplasm, nucleus and mitochondrion, at probabilities of 47.8%, 26.1% and 8.7%, respectively. In response to DNA damage NDRG1 translocates from the cytoplasm to the nucleus, where it may inhibit cell growth and promote DNA repair mechanisms. It is suggested that NDRG1 acts as a stress response gene or potentially as a transcription factor.

Background References

1. Li X et al. Golgi Phosphoprotein 3 Inhibits the Apoptosis of Human Glioma Cells in Part by Downregulating N-myc Downstream Regulated Gene 1. Med Sci Monit 22:3535-3543 (2016).

2. Lee JE & Kim JH. Valproic acid inhibits the invasion of PC3 prostate cancer cells by upregulating the metastasis suppressor protein NDRG1. Genet Mol Biol 38:527-33 (2015).

Sequence Similarity

Belongs to the NDRG family.

Tissue Specificity

Ubiquitous; expressed most prominently in placental membranes and prostate, kidney, small intestine, and ovary tissues. Also expressed in heart, brain, skeletal muscle, lung, liver and pancreas. Low levels in peripheral blood leukocytes and in tissues of the immune system. Expressed mainly in epithelial cells. Also found in Schwann cells of peripheral neurons. Reduced expression in adenocarcinomas compared to normal tissues. In colon, prostate and placental membranes, the cells that border the lumen show the highest expression.

Post-translational Modification

Under stress conditions, phosphorylated in the C-terminal on many serine and threonine residues. Phosphorylated in vitro by PKA. Phosphorylation enhanced by increased intracellular cAMP levels. Homocysteine induces dephosphorylation. Phosphorylation by SGK1 is cell cycle dependent.

Subcellular Location

Cytoplasm. Nucleus. Cell membrane.

UNIPROT

SwissProt: Q92597 Human

SwissProt: Q62433 Mouse

SwissProt: Q6JE36 Rat

Synonyms

42 kDa antibody

Anti GC4 antibody

cap43 antibody

cmt4d antibody

Differentiation related gene1 protein antibody

Differentiation-related gene 1 protein antibody

Drg 1 antibody

DRG-1 antibody

drg1 antibody

gc4 antibody

Expand

Images

  • Western blot analysis of NDRG1 on different lysates with Rabbit anti-NDRG1 antibody (<a href="/products/ET1705-20" style="font-weight: bold;text-decoration: underline;">ET1705-20</a>) at 1/1,000 dilution.<br /><br />Lane 1: PC-3 cell lysate (20 µg/Lane)<br />Lane 2: SW480 cell lysate (20 µg/Lane)<br />Lane 3: RPMI 8226 cell lysate (20 µg/Lane)<br />Lane 4: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 5: Mouse eyeball tissue lysate (40 µg/Lane)<br />Lane 6: Rat eyeball tissue lysate (40 µg/Lane)<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 43 kDa<br />Observed band size: 48 kDa<br /><br />Exposure time: 6 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/ET1705-20" style="font-weight: bold;text-decoration: underline;">ET1705-20</a>) at 1/1,000 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.

    Western blot analysis of NDRG1 on different lysates with Rabbit anti-NDRG1 antibody (ET1705-20) at 1/1,000 dilution.

    Lane 1: PC-3 cell lysate (20 µg/Lane)
    Lane 2: SW480 cell lysate (20 µg/Lane)
    Lane 3: RPMI 8226 cell lysate (20 µg/Lane)
    Lane 4: RAW264.7 cell lysate (20 µg/Lane)
    Lane 5: Mouse eyeball tissue lysate (40 µg/Lane)
    Lane 6: Rat eyeball tissue lysate (40 µg/Lane)

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 43 kDa
    Observed band size: 48 kDa

    Exposure time: 6 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 (ET1705-20) at 1/1,000 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.

  • ICC staining of NDRG1 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1705-20" style="font-weight: bold;text-decoration: underline;">ET1705-20</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of NDRG1 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1705-20, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of NDRG1 in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1705-20" style="font-weight: bold;text-decoration: underline;">ET1705-20</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of NDRG1 in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1705-20, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Immunohistochemical analysis of paraffin-embedded human prostate tissue with Rabbit anti-NDRG1 antibody (<a href="/products/ET1705-20" style="font-weight: bold;text-decoration: underline;">ET1705-20</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/ET1705-20" style="font-weight: bold;text-decoration: underline;">ET1705-20</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 prostate tissue with Rabbit anti-NDRG1 antibody (ET1705-20) 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 (ET1705-20) 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-NDRG1 antibody (<a href="/products/ET1705-20" style="font-weight: bold;text-decoration: underline;">ET1705-20</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/ET1705-20" style="font-weight: bold;text-decoration: underline;">ET1705-20</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-NDRG1 antibody (ET1705-20) 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 (ET1705-20) 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 colon carcinoma tissue using anti-NDRG1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-20" style="font-weight: bold;text-decoration: underline;">ET1705-20</a>, 1/50) for 30 minutes 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 colon carcinoma tissue using anti-NDRG1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-20, 1/50) for 30 minutes 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.

  • Application: IHC-Fr<br /><br />Species: Mouse<br /><br />Site: kidney<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/100<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Mouse

    Site: kidney

    Sample: Frozen section

    Antibody concentration: 1/100

    Antigen retrieval: Not required

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

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