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GRIM19 Recombinant Rabbit Monoclonal Antibody [JG82-33]

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Specification

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Catalog# ET7108-66

GRIM19 Recombinant Rabbit Monoclonal Antibody [JG82-33]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

GRIM19 Recombinant Rabbit Monoclonal Antibody [JG82-33]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human GRIM19 aa 1-144 / 144.

Species Reactivity

Human

Validated Applications

WB, IHC-P, IF-Cell

Target Molecular Weight

Predicted band size: 17 kDa

Positive Control

HeLa cell lysate, Ramos cell lysate, 293T cell lysate, Jurkat cell lysate, MCF7 cell lysate, human kidney tissue, human liver tissue, human prostate cancer tissue, HepG2.

Conjugation

unconjugated

Clone Number

JG82-33

RRID

AB_3073481

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

  • 1:1,000-1:2,000

  • IHC-P

  • 1:1,000

  • IF-Cell

  • 1:50-1:200

Target

Function

The GRIM family of proteins appear to be novel types of tumor suppressors. Grim19, which stands for gene associated with retinoic-interferon-induced mortality 19 protein, is also designated cell death-regulatory protein Grim-19 or NADH dehydrogenase ubiquinone 1 alpha subcomplex subunit 13. The Grim19 protein plays two roles within the cell. As a member of the interferon-beta and retinoic acid-induced pathway of cell death, Grim19 induces apoptosis. As part of the mitochondrial complex I, Grim19 is essential for its assembly and electron transfer activity. It transfers electrons to the respiratory chain from NADH and plays a role in the interferon/all-trans-retinoic acid (IFN/RA) cell death pathway. It localizes primarily to the mitochondrion, but may translocate to the nucleus upon IFN/RA treatment. Grim19 may also be useful as a biological marker or target for drug development.

Background References

1. Lufei C et al. GRIM-19, a death-regulatory gene product, suppresses Stat3 activity via functional interaction. EMBO J 22:1325-1335 (2003).

2. Zhang J et al. The cell death regulator GRIM-19 is an inhibitor of signal transducer and activator of transcription 3. Proc Natl Acad Sci USA 100:9342-9347 (2003).

Sequence Similarity

Belongs to the complex I NDUFA13 subunit family.

Tissue Specificity

Widely expressed, with highest expression in heart, skeletal muscle, liver, kidney and placenta. In intestinal mucosa, down-regulated in areas involved in Crohn disease and ulcerative colitis.

Subcellular Location

Mitochondrion. Nucleus.

UNIPROT

SwissProt: Q9P0J0 Human

Synonyms

2700054G14Rik antibody

AU022060 antibody

B16.6 antibody

CDA016 antibody

Cell death regulatory protein antibody

Cell death regulatory protein GRIM-19 antibody

CGI-39 antibody

CGI39 protein antibody

CI-B16.6 antibody

Complex I-B16.6 antibody

Expand

Images

  • Western blot analysis of GRIM19 on different lysates with Rabbit anti-GRIM19 antibody (<a href="/products/ET7108-66" style="font-weight: bold;text-decoration: underline;">ET7108-66</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: Ramos cell lysate<br />Lane 3: 293T cell lysate<br />Lane 4: Jurkat cell lysate<br />Lane 5: MCF7 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 17 kDa<br />Observed band size: 17 kDa<br /><br />Exposure time: 3 minutes;<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/ET7108-66" style="font-weight: bold;text-decoration: underline;">ET7108-66</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 GRIM19 on different lysates with Rabbit anti-GRIM19 antibody (ET7108-66) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: Ramos cell lysate
    Lane 3: 293T cell lysate
    Lane 4: Jurkat cell lysate
    Lane 5: MCF7 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 17 kDa
    Observed band size: 17 kDa

    Exposure time: 3 minutes;

    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 (ET7108-66) 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of GRIM19 on different lysates with Rabbit anti-GRIM19 antibody (<a href="/products/ET7108-66" style="font-weight: bold;text-decoration: underline;">ET7108-66</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-GRIM19 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 17 kDa<br />Observed band size: 17 kDa<br /><br />Exposure time: 60 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/ET7108-66" style="font-weight: bold;text-decoration: underline;">ET7108-66</a>) at 1/2,000 dilution was used in <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 GRIM19 on different lysates with Rabbit anti-GRIM19 antibody (ET7108-66) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-GRIM19 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 17 kDa
    Observed band size: 17 kDa

    Exposure time: 60 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 (ET7108-66) at 1/2,000 dilution was used in 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.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-GRIM19 antibody (<a href="/products/ET7108-66" style="font-weight: bold;text-decoration: underline;">ET7108-66</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/ET7108-66" style="font-weight: bold;text-decoration: underline;">ET7108-66</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-GRIM19 antibody (ET7108-66) 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 (ET7108-66) 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 liver tissue with Rabbit anti-GRIM19 antibody (<a href="/products/ET7108-66" style="font-weight: bold;text-decoration: underline;">ET7108-66</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/ET7108-66" style="font-weight: bold;text-decoration: underline;">ET7108-66</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 liver tissue with Rabbit anti-GRIM19 antibody (ET7108-66) 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 (ET7108-66) 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 cancer tissue with Rabbit anti-GRIM19 antibody (<a href="/products/ET7108-66" style="font-weight: bold;text-decoration: underline;">ET7108-66</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/ET7108-66" style="font-weight: bold;text-decoration: underline;">ET7108-66</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 cancer tissue with Rabbit anti-GRIM19 antibody (ET7108-66) 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 (ET7108-66) 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.

  • Immunocytochemistry analysis of HepG2 cells labeling GRIM19 with Rabbit anti-GRIM19 antibody (<a href="/products/ET7108-66" style="font-weight: bold;text-decoration: underline;">ET7108-66</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-GRIM19 antibody (<a href="/products/ET7108-66" style="font-weight: bold;text-decoration: underline;">ET7108-66</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of HepG2 cells labeling GRIM19 with Rabbit anti-GRIM19 antibody (ET7108-66) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-GRIM19 antibody (ET7108-66) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

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