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NUP50 Recombinant Rabbit Monoclonal Antibody [JE63-92]

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Specification

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Catalog# HA721026

NUP50 Recombinant Rabbit Monoclonal Antibody [JE63-92]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

NUP50 Recombinant Rabbit Monoclonal Antibody [JE63-92]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human NUP50 aa 331-380/468.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 50 kDa

Positive Control

HeLa (Human cervical adenocarcinoma cells) cell lysate, HCT 116 (Human colon cancer cells) cell lysate, Jurkat (Human T-lymphoblastic cells) cell lysate, Hep G2 (Human liver cancer cells) cell lysate, 293T (Human embryonic kidney cells) cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, rat testis tissue, human tonsil tissue, human thyroid tissue, human colon carcinoma tissue, human skin tissue, human breast tissue, human breast carcinoma tissue, human esophagus tissue, mouse hippocampus tissue, mouse brain tissue, Daudi, Hela.

Conjugation

unconjugated

Clone Number

JE63-92

RRID

AB_3072150

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

  • 1:2,000

  • IF-Cell

  • 1:50

  • IHC-P

  • 1:200-1:400

  • FC

  • 1:500-1:1,000

Target

Function

Nucleoporin 50 (Nup50) is a protein that in humans is encoded by the NUP50 gene. The nuclear pore complex is a massive structure that extends across the nuclear envelope, forming a gateway that regulates the flow of macromolecules between the nucleus and the cytoplasm. Nucleoporins are the main components of the nuclear pore complex in eukaryotic cells. The protein encoded by this gene is a member of the FG-repeat containing nucleoporins that functions as a soluble cofactor in importin-alpha:beta-mediated nuclear protein import. Pseudogenes of this gene are found on chromosomes 5, 6, and 14. Two transcript variants encoding different isoforms have been found for this gene.

Background References

1. Makise M. et. al. The Nup153-Nup50 protein interface and its role in nuclear import. J Biol Chem. 2012 Nov

2. Park E. et. al. NUP50 is necessary for the survival of primordial germ cells in mouse embryos. Reproduction. 2016 Jan

Subcellular Location

Nuclear pore complex, Nucleus membrane.

UNIPROT

SwissProt: Q9UKX7 Human

SwissProt: Q9JIH2 Mouse

SwissProt: O08587 Rat

Synonyms

50 kDa nucleoporin antibody

NPAP60 antibody

NPAP60L antibody

Nuclear pore associated protein 60L antibody

Nuclear pore complex protein Nup50 antibody

Nuclear pore-associated protein 60 kDa-like antibody

Nucleoporin 50 antibody

Nucleoporin 50kDa antibody

Nucleoporin Nup50 antibody

Nup50 antibody

Expand

Images

  • Western blot analysis of NUP50 on different lysates with Rabbit anti-NUP50 antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa (Human cervical adenocarcinoma cells) cell lysate<br />Lane 2: HCT 116 (Human colon cancer cells) cell lysate<br />Lane 3: Jurkat (Human T-lymphoblastic cells) cell lysate<br />Lane 4: Hep G2 (Human liver cancer cells) cell lysate<br />Lane 5: 293T (Human embryonic kidney cells) cell lysate<br />Lane 6: Mouse testis tissue lysate<br />Lane 7: Rat testis tissue lysate<br /><br />Cell Lysates/proteins at 15 µg/Lane, Tissue Lysates/proteins at 30 µg/Lane<br />Exposure time: 8 seconds; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>, 1/2,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 50 kDa<br />Observed band size: 55 kDa

    Western blot analysis of NUP50 on different lysates with Rabbit anti-NUP50 antibody (HA721026) at 1/2,000 dilution.

    Lane 1: HeLa (Human cervical adenocarcinoma cells) cell lysate
    Lane 2: HCT 116 (Human colon cancer cells) cell lysate
    Lane 3: Jurkat (Human T-lymphoblastic cells) cell lysate
    Lane 4: Hep G2 (Human liver cancer cells) cell lysate
    Lane 5: 293T (Human embryonic kidney cells) cell lysate
    Lane 6: Mouse testis tissue lysate
    Lane 7: Rat testis tissue lysate

    Cell Lysates/proteins at 15 µg/Lane, Tissue Lysates/proteins at 30 µg/Lane
    Exposure time: 8 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA721026, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 50 kDa
    Observed band size: 55 kDa

  • Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-NUP50 antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/400 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/400 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 testis tissue with Rabbit anti-NUP50 antibody (HA721026) at 1/400 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721026) at 1/400 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 tonsil tissue with Rabbit anti-NUP50 antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 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 tonsil tissue with Rabbit anti-NUP50 antibody (HA721026) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721026) at 1/200 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 thyroid tissue with Rabbit anti-NUP50 antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 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 thyroid tissue with Rabbit anti-NUP50 antibody (HA721026) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721026) at 1/200 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 with Rabbit anti-NUP50 antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 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 with Rabbit anti-NUP50 antibody (HA721026) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721026) at 1/200 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 skin tissue with Rabbit anti-NUP50 antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 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 skin tissue with Rabbit anti-NUP50 antibody (HA721026) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721026) at 1/200 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 breast tissue with Rabbit anti-NUP50 antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 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 breast tissue with Rabbit anti-NUP50 antibody (HA721026) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721026) at 1/200 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 breast carcinoma tissue with Rabbit anti-NUP50 antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 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 breast carcinoma tissue with Rabbit anti-NUP50 antibody (HA721026) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721026) at 1/200 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 esophagus tissue with Rabbit anti-NUP50 antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/200 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 esophagus tissue with Rabbit anti-NUP50 antibody (HA721026) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721026) at 1/200 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 hippocampus tissue with Rabbit anti-NUP50 antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/400 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/400 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 hippocampus tissue with Rabbit anti-NUP50 antibody (HA721026) at 1/400 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721026) at 1/400 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 brain tissue with Rabbit anti-NUP50 antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/400 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/400 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 brain tissue with Rabbit anti-NUP50 antibody (HA721026) at 1/400 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721026) at 1/400 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.

  • Flow cytometric analysis of Daudi cells labeling NUP50.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Daudi cells labeling NUP50.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721026, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Immunocytochemistry analysis of Hela cells labeling NUP50 with Rabbit anti-NUP50 antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</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-NUP50 antibody (<a href="/products/HA721026" style="font-weight: bold;text-decoration: underline;">HA721026</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor&reg; 488) 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 (Alexa Fluor&reg; 647) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of Hela cells labeling NUP50 with Rabbit anti-NUP50 antibody (HA721026) 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-NUP50 antibody (HA721026) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 (Alexa Fluor® 647) were used as the secondary antibody at 1/1,000 dilution.

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