U2AF2/U2AF65 Monoclonal antibody

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  WB analysis using 68166-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 68166-1-Ig (U2AF2 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Beta Actin Monoclonal antibody (HRP-66009) as loading control.

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  WB analysis of 4T1 using 68166-1-Ig

  4T1 cells were subjected to SDS PAGE followed by western blot with 68166-1-Ig (U2AF2 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  IHC staining of mouse liver using 68166-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse liver tissue slide using 68166-1-Ig (U2AF2 antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse liver using 68166-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse liver tissue slide using 68166-1-Ig (U2AF2 antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of rat liver using 68166-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat liver tissue slide using 68166-1-Ig (U2AF2 antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HEK-293 using 68166-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HEK-293 cells using U2AF2 antibody (68166-1-Ig, Clone: 1B7E7 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

• 

  IF Staining of HEK-293 using 68166-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HEK-293 cells using U2AF2 antibody (68166-1-Ig, Clone: 1B7E7 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

• 

  IF Staining of HEK-293 using 68166-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HEK-293 cells using U2AF2 antibody (68166-1-Ig, Clone: 1B7E7 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L), CL594-Phalloidin (red).

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  FC experiment of HEK-293 using 68166-1-Ig

  1X10^6 HEK-293 cells were intracellularly stained with 0.4 ug Anti-Human U2AF2 (68166-1-Ig, Clone:1B7E7) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG2a Isotype Control (C1.18.4) (65208-1-Ig, Clone: C1.18.4) (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

U2AF2/U2AF65 Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Cat No. 68166-1-Ig

Clone No.1B7E7

Publications(2)

Host / Isotype

Mouse / IgG2a

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Synonyms

U2AF65, U2AF2, hU2AF65, hU2AF(65), 1B7E7

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Tested Applications

• Positive WB detected in: LNCaP cells, 4T1 cells, HeLa cells, HEK-293 cells, HepG2 cells, Jurket cells, K-562 cells, HSC-T6 cells, NIH/3T3 cells
• Positive IHC detected in: mouse liver tissue, rat liver tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HEK-293 cells
• Positive FC (Intra) detected in: HEK-293 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 2 publications below

Product Information

68166-1-Ig targets U2AF2/U2AF65 in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human
• Host / Isotype: Mouse / IgG2a
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag8070

  Product name: Recombinant human U2AF2 protein

  Source: e coli.-derived, PGEX-4T

  Tag: GST

  Domain: 116-471 aa of BC008740

  Sequence: IPATALLPTMTPDGLAVTPTPVPVVGSQMTRQARRLYVGNIPFGITEEAMMDFFNAQMRLGGLTQAPGNPVLAVQINQDKNFAFLEFRSVDETTQAMAFDGIIFQGQSLKIRRPHDYQPLPGMSENPSVYVPGVVSTVVPDSAHKLFIGGLPNYLNDDQVKELLTSFGPLKAFNLVKDSATGLSKGYAFCEYVDINVTDQAIAGLNGMQLGDKKLLVQRASVGAKNATLSTINQTPVTLQVPGLMSSQVQMGGHPTEVLCLMNMVLPEELLDDEEYEEIVEDVRDECSKYGLVKSIEIPRPVDGVEVPGCGKIFVEFTSVFDCQKAMQGLTGRKFANRVVVTKYCDPDSYHRRDFW

• Full Name: U2 small nuclear RNA auxiliary factor 2
• Calculated Molecular Weight: 54 kDa
• Observed Molecular Weight: 60-65 kDa
• GenBank Accession Number: BC008740
• Gene Symbol: U2AF2
• Gene ID (NCBI): 11338
• RRID: AB_2935262
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: P26368
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

U2AF2, also named as U2 auxiliary factor 65 kDa subunit, is a 475 amino acid protein, which belongs to the splicing factor SR family. U2AF2 is Necessary for the splicing of pre-mRNA. By recruiting PRPF19 and the PRP19C/Prp19 complex/NTC/Nineteen complex to the RNA polymerase II C-terminal domain (CTD), pre-mRNA may couple transcription to splicing. The calculated molecular weight of U2AF2 is 53 kDa, but modified U2AF2 is about 65 kDa.

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
human	WB	Nat Commun; CSTF2 mediated mRNA N6-methyladenosine modification drives pancreatic ductal adenocarcinoma m6A subtypes; Authors - Yanfen Zheng
human	WB	Acta Pharm Sin B; BRD4 regulates m6A of ESPL1 mRNA via interaction with ALKBH5 to modulate breast cancer progression; Authors - Haisheng Zhang