U2AF35 Monoclonal antibody

U2AF35 Monoclonal Antibody for FC, IF, IHC, IP, WB, ELISA

Host / Isotype

Mouse / IgG2c

Reactivity

human, mouse, rat

Applications

WB, IP, IHC, IF, FC, ELISA

Conjugate

Unconjugated

Publications

4

CloneNo.

2B7F11

Cat no : 60289-1-Ig

Synonyms

FP793, RN, RNU2AF1, U2AF1, U2AF35, U2AFBP

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Validation Data Gallery

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  WB analysis using 60289-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 60289-1-Ig (U2AF35 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis of HeLa using 60289-1-Ig

  WB result of U2AF35 antibody (60289-1-Ig; 1:3000; incubated at room temperature for 1.5 hours) with sh-Control and sh-U2AF35 transfected HeLa cells.

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  WB analysis of COLO 320 using 60289-1-Ig

  COLO 320 cells were subjected to SDS PAGE followed by western blot with 60289-1-Ig (U2AF35 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 60289-1-Ig

  IP result of anti-U2AF35 (IP:60289-1-Ig, 3ug; Detection:60289-1-Ig 1:500) with HeLa cells lysate 3000ug.

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  IHC staining of human breast cancer using 60289-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 60289-1-Ig (U2AF35 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human liver cancer using 60289-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 60289-1-Ig (U2AF35 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human liver cancer using 60289-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 60289-1-Ig (U2AF35 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 60289-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 60289-1-Ig (U2AF35 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HepG2 using 60289-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using U2AF35 antibody (60289-1-Ig, Clone: 2B7F11 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  IF Staining of HEK-293 using 60289-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HEK-293 cells using U2AF35 antibody (60289-1-Ig, Clone: 2B7F11 ) at dilution of 1:2000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), CL594-phalloidin (red).

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  FC experiment of Ramos using 60289-1-Ig

  1X10^6 Ramos cells were intracellularly stained with 0.4 ug Anti-Human U2AF35 (60289-1-Ig, Clone:2B7F11) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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Tested Applications

• Positive WB detected in: HeLa cells, COLO 320 cells, HEK-293 cells, Raji cells, Ramos cells, HSC-T6 cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human breast cancer tissue, human liver cancer 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 detected in: HEK-293 cells, HepG2 cells
• Positive FC detected in: Ramos cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:6000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF): IF : 1:1000-1:4000
• Flow Cytometry (FC): FC : 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 4 publications below

Product Information

60289-1-Ig targets U2AF35 in WB, IP, IHC, IF, FC, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human
• Host / Isotype: Mouse / IgG2c
• Class: Monoclonal
• Type: Antibody
• Immunogen: U2AF35 fusion protein Ag0399
• Full Name: U2 small nuclear RNA auxiliary factor 1
• Calculated Molecular Weight: 28 kDa
• Observed Molecular Weight: 35-40 kDa
• GenBank Accession Number: BC001923
• Gene Symbol: U2AF1
• Gene ID (NCBI): 7307
• RRID: AB_2858277
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• 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

U2 auxiliary factor (U2AF), comprising a large and a small subunit, is a non-snRNP protein required for the binding of U2 snRNP to the pre-mRNA branch site. U2 (RNU2) small nuclear RNA auxiliary factor 1 (U2AF1,synonyms: RN, FP793, U2AF35, U2AFBP, RNU2AF1)is the small subunit which plays a critical role in both constitutive and enhancer-dependent RNA splicing by directly mediating interactions between the large subunit and proteins bound to the enhancers. The U2AF1 gene is localized to chromosome 21q22.3, which is the critical region for three diseases, progressive myoclonus epilepsy, autoimmune polyglandular disease type 1, and one form of bipolar affective disorder. The calculated molecular weight of U2AF35 is 28 kDa, but the modified U2AF35 protein is about 35-40 kDa.

Publications

Species	Application	Title
human	WB	Nat Commun; SPF45/RBM17-dependent, but not U2AF-dependent, splicing in a distinct subset of human short introns.; Authors - Kazuhiro Fukumura
human	WB	Mol Neurodegener; Nuclear speckle specific hnRNP D-like prevents age- and AD-related cognitive decline by modulating RNA splicing.; Authors - Qingyang Zhang
human	WB	Nucleic Acids Res; Stoichiometries of U2AF35, U2AF65 and U2 snRNP reveal new early spliceosome assembly pathways.; Authors - Li Chen
human	WB	RNA; A model for DHX15 mediated disassembly of A-complex spliceosomes; Authors - Hannah M Maul-Newby