U2AF65 Recombinant Rabbit Monoclonal Antibody [JE36-33]
Catalog# HA721466
U2AF65 Recombinant Rabbit Monoclonal Antibody [JE36-33]
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WB
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IHC-P
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Human
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Mouse
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Rat
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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
Target 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
Product Features
Form
Liquid
Concentration
1 mg/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
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1:1,000
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IHC-P
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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.
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
ExpandImages
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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 (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 (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 (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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"