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Aly Recombinant Rabbit Monoclonal Antibody [JE36-45]

Usd: 385

Specification

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Catalog# HA721961

Aly Recombinant Rabbit Monoclonal Antibody [JE36-45]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

Aly Recombinant Rabbit Monoclonal Antibody [JE36-45]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Aly aa 208-257 / 257.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 27 kDa

Positive Control

HepG2 cell lysate, K-562 cell lysate, 293T cell lysate, A549 cell lysate, SK-OV-3 cell lysate, HL-60 cell lysate, NIH/3T3 cell lysate, Neuro-2a cell lysate, C6 cell lysate, mouse testis tissue lysate, mouse spleen tissue lysate, rat testis tissue lysate, HepG2, human brain tissue, mouse brain tissue, rat brain tissue.

Conjugation

unconjugated

Clone Number

JE36-45

RRID

AB_3096740

Product Features

Form

Liquid

Concentration

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

  • 1:250

  • IHC-P

  • 1:2,000

  • FC

  • 1:1,000

Target

Function

Aly/REF export factor, also known as THO complex subunit 4 is a protein that in humans is encoded by the ALYREF gene. The ALYREF gene encodes Aly/REF export factor (ALY; THO complex subunit 4, Tho4; RNA and export factor binding protein 1, Refbp1), a ubiquitously expressed nuclear protein that functions as a molecular chaperone and export adapter involved in nuclear export of spliced and unspliced mRNA. The TRanscription-EXport (TREX) complex, a key player in mRNA export, includes the THO subcomplex, the RNA helicase UAP56, and the RNA-binding protein ALY. In yeast, TREX is recruited co-transcriptionally; in human cells it is recruited during a late step of splicing. The human TREX complex is recruited to a region near the 5' end of mRNA by interaction of ALY and THO with the nuclear cap-binding complex. As a chaperone, ALY promotes dimerization of transcription factors containing basic leucine zipper (bZIP) domains., thereby promoting transcriptional activation. ALY has key roles in 3'-end processing of polyadenylated mRNAs and in nuclear export of both polyadenylated and non-polyadenylated mRNAs. After mRNA binds to ALY, it is apparently transferred to the NXF1-NXT1 heterodimer for export (TAP/NFX1 pathway). The full-length ALY protein (Refbp1-I, 255 amino acids encoded by six exons[8]) has a conserved RNA recognition motif (RRM; amino acids 105-182) flanked by alanine/arginine/glycine-rich sequences; an N-terminal region (amino acids 16-37) is sufficient for RNA binding and interaction with the NXF1-NXT1 heterodimer.

Background References

1. Kow RL et al. Loss of aly/ALYREF suppresses toxicity in both tau and TDP-43 models of neurodegeneration. Geroscience. 2022 Apr

2. Wang JZ et al. The role of the HIF-1α/ALYREF/PKM2 axis in glycolysis and tumorigenesis of bladder cancer. Cancer Commun (Lond). 2021 Jul

Subcellular Location

Nucleus, Nucleus speckle, Cytoplasm.

UNIPROT

SwissProt: Q86V81 Human

SwissProt: O08583 Mouse

Entrez Gene: 690585 Rat

Synonyms

Ally of AML-1 and LEF-1 antibody

Ally of AML1 and LEF1 antibody

ALY antibody

ALY/REF antibody

Aly/REF export factor antibody

BEF antibody

bZIP enhancing factor antibody

bZIP-enhancing factor BEF antibody

REF antibody

THO complex 4 antibody

Expand

Images

  • Western blot analysis of Aly on different lysates with Rabbit anti-Aly antibody (<a href="/products/HA721961" style="font-weight: bold;text-decoration: underline;">HA721961</a>) at 1/1,000 dilution.<br /><br />Lane 1: HepG2 cell lysate (20 µg/Lane)<br />Lane 2: K-562 cell lysate (20 µg/Lane)<br />Lane 3: 293T cell lysate (20 µg/Lane)<br />Lane 4: A549 cell lysate (20 µg/Lane)<br />Lane 5: SK-OV-3 cell lysate (20 µg/Lane)<br />Lane 6: HL-60 cell lysate (20 µg/Lane)<br />Lane 7: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 8: Neuro-2a cell lysate (20 µg/Lane)<br />Lane 9: C6 cell lysate (20 µg/Lane)<br />Lane 10: Mouse testis tissue lysate (40 µg/Lane)<br />Lane 11: Mouse spleen tissue lysate (40 µg/Lane)<br />Lane 12: Rat testis tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 27 kDa<br />Observed band size: 27 kDa<br /><br />Exposure time: 2 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/HA721961" style="font-weight: bold;text-decoration: underline;">HA721961</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 Aly on different lysates with Rabbit anti-Aly antibody (HA721961) at 1/1,000 dilution.

    Lane 1: HepG2 cell lysate (20 µg/Lane)
    Lane 2: K-562 cell lysate (20 µg/Lane)
    Lane 3: 293T cell lysate (20 µg/Lane)
    Lane 4: A549 cell lysate (20 µg/Lane)
    Lane 5: SK-OV-3 cell lysate (20 µg/Lane)
    Lane 6: HL-60 cell lysate (20 µg/Lane)
    Lane 7: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 8: Neuro-2a cell lysate (20 µg/Lane)
    Lane 9: C6 cell lysate (20 µg/Lane)
    Lane 10: Mouse testis tissue lysate (40 µg/Lane)
    Lane 11: Mouse spleen tissue lysate (40 µg/Lane)
    Lane 12: Rat testis tissue lysate (40 µg/Lane)

    Predicted band size: 27 kDa
    Observed band size: 27 kDa

    Exposure time: 2 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 (HA721961) 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.

  • Immunocytochemistry analysis of HepG2 cells labeling Aly with Rabbit anti-Aly antibody (<a href="/products/HA721961" style="font-weight: bold;text-decoration: underline;">HA721961</a>) at 1/250 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 Rabbit anti-Aly antibody (<a href="/products/HA721961" style="font-weight: bold;text-decoration: underline;">HA721961</a>) at 1/250 dilution in 1% BSA in PBST 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. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of HepG2 cells labeling Aly with Rabbit anti-Aly antibody (HA721961) at 1/250 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 Rabbit anti-Aly antibody (HA721961) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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.

  • Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-Aly antibody (<a href="/products/HA721961" style="font-weight: bold;text-decoration: underline;">HA721961</a>) at 1/2,000 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/HA721961" style="font-weight: bold;text-decoration: underline;">HA721961</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 human brain tissue with Rabbit anti-Aly antibody (HA721961) at 1/2,000 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 (HA721961) 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 brain tissue with Rabbit anti-Aly antibody (<a href="/products/HA721961" style="font-weight: bold;text-decoration: underline;">HA721961</a>) at 1/2,000 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/HA721961" style="font-weight: bold;text-decoration: underline;">HA721961</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 brain tissue with Rabbit anti-Aly antibody (HA721961) at 1/2,000 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 (HA721961) 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 rat brain tissue with Rabbit anti-Aly antibody (<a href="/products/HA721961" style="font-weight: bold;text-decoration: underline;">HA721961</a>) at 1/2,000 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/HA721961" style="font-weight: bold;text-decoration: underline;">HA721961</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 rat brain tissue with Rabbit anti-Aly antibody (HA721961) at 1/2,000 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 (HA721961) 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.

  • Flow cytometric analysis of HepG2 cells labeling Aly.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721961" style="font-weight: bold;text-decoration: underline;">HA721961</a>, 1μg/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 HepG2 cells labeling Aly.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721961, 1μg/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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