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NMI Recombinant Rabbit Monoclonal Antibody [JE65-78]

Usd: 385

Specification

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Catalog# HA720087

NMI Recombinant Rabbit Monoclonal Antibody [JE65-78]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

NMI Recombinant Rabbit Monoclonal Antibody [JE65-78]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human NMI aa 1-51/307.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IHC-P, FC

Target Molecular Weight

Predicted band size: 35 kDa

Positive Control

THP-1 cell lysate, HeLa cell lysate, K-562 cell lysate, human spleen tissue, Hela, SiHa.

Conjugation

unconjugated

Clone Number

JE65-78

RRID

AB_3072026

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

  • IF-Cell

  • 1:50

  • IHC-P

  • 1:400

  • FC

  • 1:500-1:1,000

Target

Function

N-myc-interactor also known as N-myc and STAT interactor is a protein that in humans is encoded by the NMI gene. NMYC interactor (NMI) encodes a protein that interacts with NMYC and CMYC (two members of the oncogene Myc family), and other transcription factors containing a Zip, HLH, or HLH-Zip motif. The NMI protein also interacts with all STATs except STAT2 and augments STAT-mediated transcription in response to cytokines IL2 and IFN-gamma. The NMI mRNA has low expression levels in all human fetal and adult tissues tested except brain and has high expression in cancer cell line-myeloid leukemias.

Background References

1. Ji D. et. al. NMI promotes cell proliferation through TGFbeta/Smad pathway by upregulating STAT1 in colorectal cancer. J Cell Physiol. 2020 Jan

2. Xu X. et. al. Interferon activates promoter of Nmi gene via interferon regulator factor-1. Mol Cell Biochem. 2018 Apr

Subcellular Location

Nucleus, Secreted, Cytoplasm.

UNIPROT

SwissProt: Q13287 Human

Synonyms

N myc (and STAT) interactor antibody

N myc and STAT interactor antibody

N myc interactor antibody

NMI antibody

NMYC and STAT interactor antibody

NMYC interactor antibody

Images

  • Western blot analysis of NMI on different lysates with Rabbit anti-NMI antibody (<a href="/products/HA720087" style="font-weight: bold;text-decoration: underline;">HA720087</a>) at 1/5,000 dilution.<br /><br />Lane 1: THP-1 cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (20 µg/Lane)<br />Lane 3: K-562 cell lysate (20 µg/Lane)<br /><br />Predicted band size: 35 kDa<br />Observed band size: 39 kDa<br />Exposure time: 25 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/HA720087" style="font-weight: bold;text-decoration: underline;">HA720087</a>) at 1/5,000 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.

    Western blot analysis of NMI on different lysates with Rabbit anti-NMI antibody (HA720087) at 1/5,000 dilution.

    Lane 1: THP-1 cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (20 µg/Lane)
    Lane 3: K-562 cell lysate (20 µg/Lane)

    Predicted band size: 35 kDa
    Observed band size: 39 kDa
    Exposure time: 25 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 (HA720087) at 1/5,000 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.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-NMI antibody (<a href="/products/HA720087" style="font-weight: bold;text-decoration: underline;">HA720087</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/HA720087" style="font-weight: bold;text-decoration: underline;">HA720087</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 spleen tissue with Rabbit anti-NMI antibody (HA720087) 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 (HA720087) 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 Hela cells labeling NMI with Rabbit anti-NMI antibody (<a href="/products/HA720087" style="font-weight: bold;text-decoration: underline;">HA720087</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-NMI antibody (<a href="/products/HA720087" style="font-weight: bold;text-decoration: underline;">HA720087</a>) at 1/50 dilution in 2% negative goat serum 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. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 647, <a href="/products/HA1127" style="font-weight: bold;text-decoration: underline;">HA1127</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of Hela cells labeling NMI with Rabbit anti-NMI antibody (HA720087) 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-NMI antibody (HA720087) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 647, HA1127) were used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of SiHa cells labeling NMI with Rabbit anti-NMI antibody (<a href="/products/HA720087" style="font-weight: bold;text-decoration: underline;">HA720087</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-NMI antibody (<a href="/products/HA720087" style="font-weight: bold;text-decoration: underline;">HA720087</a>) at 1/50 dilution in 2% negative goat serum 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. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 647, <a href="/products/HA1127" style="font-weight: bold;text-decoration: underline;">HA1127</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of SiHa cells labeling NMI with Rabbit anti-NMI antibody (HA720087) 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-NMI antibody (HA720087) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 647, HA1127) were used as the secondary antibody at 1/1,000 dilution.

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