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NM23 Recombinant Mouse Monoclonal Antibody [13C2-R]

Usd: 350

Specification

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Catalog# HA601307

NM23 Recombinant Mouse Monoclonal Antibody [13C2-R]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

NM23 Recombinant Mouse Monoclonal Antibody [13C2-R]

Antibody Type

Recombinant Mouse Monoclonal Antibody

Immunogen

Synthetic peptide within Human NM23 aa 1-50 / 152.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell

Molecular Weight

Predicted band size: 17 kDa

Positive Control

HEK-293 cell lysate, MCF7 cell lysate, Jurkat cell lysate, Raji cell lysate, HeLa cell lysate, K-562 cell lysate, A549 cell lysate, A431 cell lysate, HepG2 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, mouse kidney tissue lysate, mouse liver tissue lysate, rat brain tissue lysate, mouse kidney tissue, HeLa, NIH/3T3, C6.

Conjugation

unconjugated

Clone Number

13C2-R

RRID

AB_3096803

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

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:2,000

  • IF-Cell

  • 1:100

Target

Function

Major role in the synthesis of nucleoside triphosphates other than ATP. The ATP gamma phosphate is transferred to the NDP beta phosphate via a ping-pong mechanism, using a phosphorylated active-site intermediate. Possesses nucleoside-diphosphate kinase, serine/threonine-specific protein kinase, geranyl and farnesyl pyrophosphate kinase, histidine protein kinase and 3'-5' exonuclease activities. Involved in cell proliferation, differentiation and development, signal transduction, G protein-coupled receptor endocytosis, and gene expression. Required for neural development including neural patterning and cell fate determination. During GZMA-mediated cell death, works in concert with TREX1. NME1 nicks one strand of DNA and TREX1 removes bases from the free 3' end to enhance DNA damage and prevent DNA end reannealing and rapid repair.

Background References

1. Fan Z. et. al. Tumor suppressor NM23-H1 is a granzyme A-activated DNase during CTL-mediated apoptosis, and the nucleosome assembly protein SET is its inhibitor. Cell 112:659-672(2003).

2. Chowdhury D. et. al. The exonuclease TREX1 is in the SET complex and acts in concert with NM23-H1 to degrade DNA during granzyme A-mediated cell death. Mol. Cell 23:133-142(2006).

Subcellular Location

Nucleus, Cytoplasm.

UNIPROT

SwissProt: P15531 Human

SwissProt: P15532 Mouse

SwissProt: Q05982 Rat

Synonyms

AWD antibody

AWD, drosophila, homolog of antibody

GAAD antibody

Granzyme A activated DNase antibody

Granzyme A-activated DNase antibody

GZMA activated DNase antibody

Metastasis inhibition factor NM23 antibody

NB antibody

NBS antibody

NDK A antibody

Expand

Images

  • Western blot analysis of NM23 on different lysates with Mouse anti-NM23 antibody (<a href="/products/HA601307" style="font-weight: bold;text-decoration: underline;">HA601307</a>) at 1/2,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate (20 µg/Lane)<br />Lane 2: MCF7 cell lysate (20 µg/Lane)<br />Lane 3: Jurkat cell lysate (20 µg/Lane)<br />Lane 4: Raji cell lysate (20 µg/Lane)<br />Lane 5: HeLa cell lysate (20 µg/Lane)<br />Lane 6: K-562 cell lysate (20 µg/Lane)<br />Lane 7: A549 cell lysate (20 µg/Lane)<br />Lane 8: A431 cell lysate (20 µg/Lane)<br />Lane 9: HepG2 cell lysate (20 µg/Lane)<br />Lane 10: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 11: C6 cell lysate (20 µg/Lane)<br />Lane 12: Mouse kidney tissue lysate (40 µg/Lane)<br />Lane 13: Mouse liver tissue lysate (40 µg/Lane)<br />Lane 14: Rat brain tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 17 kDa<br />Observed band size: 17/20 kDa<br /><br />Exposure time: 1 minute 59 seconds;<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/HA601307" style="font-weight: bold;text-decoration: underline;">HA601307</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of NM23 on different lysates with Mouse anti-NM23 antibody (HA601307) at 1/2,000 dilution.

    Lane 1: HEK-293 cell lysate (20 µg/Lane)
    Lane 2: MCF7 cell lysate (20 µg/Lane)
    Lane 3: Jurkat cell lysate (20 µg/Lane)
    Lane 4: Raji cell lysate (20 µg/Lane)
    Lane 5: HeLa cell lysate (20 µg/Lane)
    Lane 6: K-562 cell lysate (20 µg/Lane)
    Lane 7: A549 cell lysate (20 µg/Lane)
    Lane 8: A431 cell lysate (20 µg/Lane)
    Lane 9: HepG2 cell lysate (20 µg/Lane)
    Lane 10: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 11: C6 cell lysate (20 µg/Lane)
    Lane 12: Mouse kidney tissue lysate (40 µg/Lane)
    Lane 13: Mouse liver tissue lysate (40 µg/Lane)
    Lane 14: Rat brain tissue lysate (40 µg/Lane)

    Predicted band size: 17 kDa
    Observed band size: 17/20 kDa

    Exposure time: 1 minute 59 seconds;

    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 (HA601307) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Mouse anti-NM23 antibody (<a href="/products/HA601307" style="font-weight: bold;text-decoration: underline;">HA601307</a>) at 1/2,000 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/HA601307" style="font-weight: bold;text-decoration: underline;">HA601307</a>) at 1/2,000 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 Mouse anti-NM23 antibody (HA601307) at 1/2,000 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 (HA601307) at 1/2,000 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 NM23 with Mouse anti-NM23 antibody (<a href="/products/HA601307" style="font-weight: bold;text-decoration: underline;">HA601307</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 Mouse anti-NM23 antibody (<a href="/products/HA601307" style="font-weight: bold;text-decoration: underline;">HA601307</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</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/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling NM23 with Mouse anti-NM23 antibody (HA601307) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 Mouse anti-NM23 antibody (HA601307) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling NM23 with Mouse anti-NM23 antibody (<a href="/products/HA601307" style="font-weight: bold;text-decoration: underline;">HA601307</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 Mouse anti-NM23 antibody (<a href="/products/HA601307" style="font-weight: bold;text-decoration: underline;">HA601307</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</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/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of NIH/3T3 cells labeling NM23 with Mouse anti-NM23 antibody (HA601307) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 Mouse anti-NM23 antibody (HA601307) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of C6 cells labeling NM23 with Mouse anti-NM23 antibody (<a href="/products/HA601307" style="font-weight: bold;text-decoration: underline;">HA601307</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 Mouse anti-NM23 antibody (<a href="/products/HA601307" style="font-weight: bold;text-decoration: underline;">HA601307</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</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/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of C6 cells labeling NM23 with Mouse anti-NM23 antibody (HA601307) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 Mouse anti-NM23 antibody (HA601307) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of NM23 on different lysates with Mouse anti-NM23 antibody (<a href="/products/HA601307" style="font-weight: bold;text-decoration: underline;">HA601307</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549-si NT cell lysate<br />Lane 2: A549-si NM23 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 17 kDa<br />Observed band size: 17/20 kDa<br /><br />Exposure time: 30 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/HA601307" style="font-weight: bold;text-decoration: underline;">HA601307</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of NM23 on different lysates with Mouse anti-NM23 antibody (HA601307) at 1/2,000 dilution.

    Lane 1: A549-si NT cell lysate
    Lane 2: A549-si NM23 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 17 kDa
    Observed band size: 17/20 kDa

    Exposure time: 30 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 (HA601307) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

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

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