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U2AF65 Recombinant Rabbit Monoclonal Antibody [JE36-33]

Usd: 385

Specification

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Catalog# HA721466

U2AF65 Recombinant Rabbit Monoclonal Antibody [JE36-33]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

U2AF65 Recombinant Rabbit Monoclonal Antibody [JE36-33]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human U2AF65 aa 376-475 / 475.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 54 kDa

Positive Control

HEK-293 cell lysate, HepG2 cell lysate, HeLa cell lysate, MCF7 cell lysate, SH-SY5Y cell lysate, RAW264.7 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, human liver tissue, mouse liver tissue, rat brain tissue.

Conjugation

unconjugated

Clone Number

JE36-33

RRID

AB_3072582

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

  • IHC-P

  • 1:1,000

Target

Function

Splicing factor U2AF 65 kDa subunit is a protein that in humans is encoded by the U2AF2 gene. In eukaryotes, the introns in the transcribed pre-mRNA first have to be removed by spliceosome in order to form a mature mRNA. A spliceosome is assembled from small nuclear ribonucleoproteins(snRNP) and small nuclear RNAs(snRNA). And the splicing factor can be divided into snRNP and non snRNP proteins.U2 auxiliary factor (U2AF), composed of a large and a small subunit, is a non-snRNP protein required for the binding of U2 snRNP to the pre-mRNA branch site. This gene encodes the U2AF large subunit, which contains a sequence-specific RNA-binding region with 3 RNA recognition motifs and an Arg/Ser-rich domain necessary for splicing. The large subunit binds to the polypyrimidine tract of introns early during spliceosome assembly. Multiple alternatively spliced transcript variants have been detected for this gene, but the full-length natures of only two have been determined to date. In humans and other tetrapods, it has been shown that without U2AF2, the splicing process is inhibited. However, in zebrafish and other teleosts the RNA splicing process can still occur on certain genes in the absence of U2AF2. This may be because 10% of genes have alternating TG and AC base pairs at the 3' splice site (3'ss) and 5' splice site (5'ss) respectively on each intron, which alters the secondary structure of the RNA and influences splicing. The splicing factor U2AF65 can specifically recognizes the polypyrimidine tract (Py tract), that’s because U2AF65 consists of 3 RNA binding domains (RRMs), all of them have a high binding affinity to the Py tract on its adjacent 3’ splice site.[6] The RRM1 and RRM2 are sufficient for specific RNA/protein binding, while RRM3 is responsible for protein/protein interactions. For example, the C-Terminal RRM3 contribute to establish protein–protein contacts with splicing factors like UAP56, SAP155, and mBBP/SF1.

Background References

1. Choi N et al. U2AF65-Dependent SF3B1 Function in SMN Alternative Splicing. Cells. 2020 Dec

2. Tong D. The role of JMJD6/U2AF65/AR-V7 axis in castration-resistant prostate cancer progression. Cancer Cell Int. 2021 Jan

Subcellular Location

Nucleus.

UNIPROT

SwissProt: P26368 Human

SwissProt: P26369 Mouse

Entrez Gene: 308335 Rat

Synonyms

hU2AF(65) antibody

hU2AF65 antibody

Splicing factor U2AF 65 kDa subunit antibody

U2 (RNU2) small nuclear RNA auxiliary factor 2 antibody

U2 auxiliary factor 65 kDa subunit antibody

U2 small nuclear ribonucleoprotein auxiliary factor (65kD) antibody

U2 small nuclear RNA auxiliary factor 2 antibody

U2 snRNP auxiliary factor large subunit antibody

U2af2 antibody

U2AF2_HUMAN antibody

Expand

Images

  • Western blot analysis of U2AF65 on different lysates with Rabbit anti-U2AF65 antibody (<a href="/products/HA721466" style="font-weight: bold;text-decoration: underline;">HA721466</a>) at 1/1,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate<br />Lane 2: HepG2 cell lysate<br />Lane 3: HeLa cell lysate<br />Lane 4: MCF7 cell lysate<br />Lane 5: SH-SY5Y cell lysate<br />Lane 6: RAW264.7 cell lysate<br />Lane 7: NIH/3T3 cell lysate<br />Lane 8: PC-12 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 54 kDa<br />Observed band size: 54 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/HA721466" style="font-weight: bold;text-decoration: underline;">HA721466</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 U2AF65 on different lysates with Rabbit anti-U2AF65 antibody (HA721466) at 1/1,000 dilution.

    Lane 1: HEK-293 cell lysate
    Lane 2: HepG2 cell lysate
    Lane 3: HeLa cell lysate
    Lane 4: MCF7 cell lysate
    Lane 5: SH-SY5Y cell lysate
    Lane 6: RAW264.7 cell lysate
    Lane 7: NIH/3T3 cell lysate
    Lane 8: PC-12 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 54 kDa
    Observed band size: 54 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 (HA721466) 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 liver tissue with Rabbit anti-U2AF65 antibody (<a href="/products/HA721466" style="font-weight: bold;text-decoration: underline;">HA721466</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/HA721466" style="font-weight: bold;text-decoration: underline;">HA721466</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 liver tissue with Rabbit anti-U2AF65 antibody (HA721466) 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 (HA721466) 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 mouse liver tissue with Rabbit anti-U2AF65 antibody (<a href="/products/HA721466" style="font-weight: bold;text-decoration: underline;">HA721466</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/HA721466" style="font-weight: bold;text-decoration: underline;">HA721466</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 mouse liver tissue with Rabbit anti-U2AF65 antibody (HA721466) 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 (HA721466) 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 rat brain tissue with Rabbit anti-U2AF65 antibody (<a href="/products/HA721466" style="font-weight: bold;text-decoration: underline;">HA721466</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/HA721466" style="font-weight: bold;text-decoration: underline;">HA721466</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 rat brain tissue with Rabbit anti-U2AF65 antibody (HA721466) 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 (HA721466) 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.

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