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NFE2 Recombinant Rabbit Monoclonal Antibody [JE65-92]

Usd: 385

Specification

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Catalog# HA721365

NFE2 Recombinant Rabbit Monoclonal Antibody [JE65-92]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

NFE2 Recombinant Rabbit Monoclonal Antibody [JE65-92]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human NFE2 aa 251-350 / 373.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell, FC

Target Molecular Weight

Predicted band size: 41 kDa

Positive Control

U-937 cell lysate, K-562 cell lysate, TF-1 cell lysate, HeLa cell lysate, HepG2 cell lysate, mouse testis tissue lysate, rat testis tissue lysate, human breast carcinoma tissue, SH-SY5Y, K-562.

Conjugation

unconjugated

Clone Number

JE65-92

RRID

AB_3072482

Product Features

Form

Liquid

Concentration

1mg/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

  • 1:1000

  • IHC-P

  • 1:200-1:1000

  • IF-Cell

  • 1:100

  • FC

  • 1:1000

Target

Function

Component of the NF-E2 complex essential for regulating erythroid and megakaryocytic maturation and differentiation. Binds to the hypersensitive site 2 (HS2) of the beta-globin control region (LCR). This subunit (NFE2) recognizes the TCAT/C sequence of the AP-1-like core palindrome present in a number of erythroid and megakaryocytic gene promoters. Requires MAFK or other small MAF proteins for binding to the NF-E2 motif. May play a role in all aspects of hemoglobin production from globin and heme synthesis to procurement of iron.

Background References

1. H L Hung et al. Stimulation of NF-E2 DNA binding by CREB-binding protein (CBP)-mediated acetylation. J Biol Chem 276(14):10715-21 (2001)

2. Yu-Chiau Shyu et al. Sumoylation of p45/NF-E2: nuclear positioning and transcriptional activation of the mammalian beta-like globin gene locus. Mol Cell Biol 25(23):10365-78 (2005)

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: Q16621 Human

SwissProt: Q07279 Mouse

SwissProt: Q6AYT2 Rat

Synonyms

Leucine zipper protein NF-E2

Nuclear factor, erythroid-derived 2 45 kDa subunit

p45 NF-E2

Images

  • Western blot analysis of NFE2 on different lysates with Rabbit anti-NFE2 antibody (<a href="/products/HA721365" style="font-weight: bold;text-decoration: underline;">HA721365</a>) at 1/1,000 dilution.<br /><br />Lane 1: U-937 cell lysate (15 µg/Lane)<br />Lane 2: K-562 cell lysate (15 µg/Lane)<br />Lane 3: TF-1 cell lysate (15 µg/Lane)<br />Lane 4: HeLa cell lysate (15 µg/Lane)<br />Lane 5: HepG2 cell lysate (15 µg/Lane)<br />Lane 6: Mouse testis tissue lysate (30µg/Lane)<br />Lane 7: Rat testis tissue lysate (30µg/Lane)<br /><br />Predicted band size: 41 kDa<br />Observed band size: 41 kDa<br /><br />Exposure time: 1 minute 20 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/HA721365" style="font-weight: bold;text-decoration: underline;">HA721365</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of NFE2 on different lysates with Rabbit anti-NFE2 antibody (HA721365) at 1/1,000 dilution.

    Lane 1: U-937 cell lysate (15 µg/Lane)
    Lane 2: K-562 cell lysate (15 µg/Lane)
    Lane 3: TF-1 cell lysate (15 µg/Lane)
    Lane 4: HeLa cell lysate (15 µg/Lane)
    Lane 5: HepG2 cell lysate (15 µg/Lane)
    Lane 6: Mouse testis tissue lysate (30µg/Lane)
    Lane 7: Rat testis tissue lysate (30µg/Lane)

    Predicted band size: 41 kDa
    Observed band size: 41 kDa

    Exposure time: 1 minute 20 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 (HA721365) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-NFE2 antibody (<a href="/products/HA721365" style="font-weight: bold;text-decoration: underline;">HA721365</a>) at 1/1,000 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/HA721365" style="font-weight: bold;text-decoration: underline;">HA721365</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 colon cancer tissue with Rabbit anti-NFE2 antibody (HA721365) at 1/1,000 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 (HA721365) 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 testis tissue with Rabbit anti-NFE2 antibody (<a href="/products/HA721365" style="font-weight: bold;text-decoration: underline;">HA721365</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/HA721365" style="font-weight: bold;text-decoration: underline;">HA721365</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 testis tissue with Rabbit anti-NFE2 antibody (HA721365) 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 (HA721365) 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-NFE2 antibody (<a href="/products/HA721365" style="font-weight: bold;text-decoration: underline;">HA721365</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/HA721365" style="font-weight: bold;text-decoration: underline;">HA721365</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-NFE2 antibody (HA721365) 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 (HA721365) 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.

  • Immunocytochemistry analysis of K-562 cells labeling NFE2 with Rabbit anti-NFE2 antibody (<a href="/products/HA721365" style="font-weight: bold;text-decoration: underline;">HA721365</a>) at 1/100 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-NFE2 antibody (<a href="/products/HA721365" style="font-weight: bold;text-decoration: underline;">HA721365</a>) at 1/100 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; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of K-562 cells labeling NFE2 with Rabbit anti-NFE2 antibody (HA721365) at 1/100 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-NFE2 antibody (HA721365) at 1/100 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™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of K-562 cells labeling NFE2.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721365" style="font-weight: bold;text-decoration: underline;">HA721365</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 K-562 cells labeling NFE2.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721365, 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).

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