Recombinant Anti-SF2 antibody [EPR8239] (ab129108)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR8239] to SF2
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SF2 antibody [EPR8239]
See all SF2 primary antibodies -
Description
Rabbit monoclonal [EPR8239] to SF2 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HepG2, Raji, HeLa, 293T cell lysates; mouse spleen. IHC-P: Human prostatic hyperplasia and breast carcinoma, rat kidney and mouse cerebral cortex tissues. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 8.00 x 10 -12 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR8239 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab129108 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/10000. Detects a band of approximately 22-32 kDa (predicted molecular weight: 28 kDa).
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IHC-P |
1/800 - 1/1500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
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ICC/IF | (1) |
1/150 - 1/500.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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WB
1/10000. Detects a band of approximately 22-32 kDa (predicted molecular weight: 28 kDa). |
IHC-P
1/800 - 1/1500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
ICC/IF
1/150 - 1/500. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
Target
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Function
Plays a role in preventing exon skipping, ensuring the accuracy of splicing and regulating alternative splicing. Interacts with other spliceosomal components, via the RS domains, to form a bridge between the 5'- and 3'-splice site binding components, U1 snRNP and U2AF. Can stimulate binding of U1 snRNP to a 5'-splice site-containing pre-mRNA. Binds to purine-rich RNA sequences, either the octamer, 5'-RGAAGAAC-3' (r=A or G) or the decamers, AGGACAGAGC/AGGACGAAGC. Binds preferentially to the 5'-CGAGGCG-3' motif in vitro. Three copies of the octamer constitute a powerful splicing enhancer in vitro, the ASF/SF2 splicing enhancer (ASE) which can specifically activate ASE-dependent splicing. Isoform ASF-2 and isoform ASF-3 act as splicing repressors. -
Sequence similarities
Belongs to the splicing factor SR family.
Contains 2 RRM (RNA recognition motif) domains. -
Domain
The RRM 2 domain plays an important role in governing both the binding mode and the phosphorylation mechanism of the RS domain by SRPK1. RS domain and RRM 2 are uniquely positioned to initiate a highly directional (C-terminus to N-terminus) phosphorylation reaction in which the RS domain slides through an extended electronegative channel separating the docking groove of SRPK1 and the active site. RRM 2 binds toward the periphery of the active site and guides the directional phosphorylation mechanism. Both the RS domain and an RRM domain are required for nucleocytoplasmic shuttling. -
Post-translational
modificationsPhosphorylated by CLK1, CLK2, CLK3 and CLK4. Phosphorylated by SRPK1 at multiple serines in its RS domain via a directional (C-terminal to N-terminal) and a dual-track mechanism incorporating both processive phosphorylation (in which the kinase stays attached to the substrate after each round of phosphorylation) and distributive phosphorylation steps (in which the kinase and substrate dissociate after each phosphorylation event). The RS domain of SRSF1 binds to a docking groove in the large lobe of the kinase domain of SRPK1 and this induces certain structural changes in SRPK1 and/or RRM 2 domain of SRSF1, allowing RRM 2 to bind the kinase and initiate phosphorylation. The cycles continue for several phosphorylation steps in a processive manner (steps 1-8) until the last few phosphorylation steps (approximately steps 9-12). During that time, a mechanical stress induces the unfolding of the beta-4 motif in RRM 2, which then docks at the docking groove of SRPK1. This also signals RRM 2 to begin to dissociate, which facilitates SRSF1 dissociation after phosphorylation is completed.
Arg-97 is dimethylated, probably to asymmetric dimethylarginine. -
Cellular localization
Cytoplasm. Nucleus speckle. In nuclear speckles. Shuttles between the nucleus and the cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 6426 Human
- Entrez Gene: 110809 Mouse
- Entrez Gene: 689890 Rat
- Omim: 600812 Human
- SwissProt: Q07955 Human
- SwissProt: Q6PDM2 Mouse
- Unigene: 68714 Human
- Unigene: 391719 Mouse
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Alternative names
- Alternative splicing factor 1 antibody
- Alternative-splicing factor 1 antibody
- arginine/serine-rich 1 antibody
see all
Images
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Anti-SF2 antibody [EPR8239] (ab129108) at 1/10000 dilution (purified) + Mouse spleen tissue at 20 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 28 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic hyperplasia tissue labelling SF2 with purified ab129108 at 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Intracellular Flow Cytometry analysis of HeLa cells labelling SF2 with purified ab129108 at a dilution of 1/70 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Flour® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling SF2 with purified ab129108 at 1/150. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Counterstained with DAPI.
Control - primary antibody (1/150), secondary antibody ab150120 an Alexa Fluor® 594-conjugate goat anti-mouse IgG (1/500).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling SF2 with purified ab129108 at 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebral cortex tissue labelling SF2 with purified ab129108 at 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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All lanes : Anti-SF2 antibody [EPR8239] (ab129108) at 1/10000 dilution (purified)
Lane 1 : HepG2 cell lysate
Lane 2 : Raji cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : 293T cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 28 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling SF2 with unpurified ab129108 at 1/100.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab129108 has been referenced in 10 publications.
- Woodcock CC et al. Matrix stiffening induces a pathogenic QKI-miR-7-SRSF1 signaling axis in pulmonary arterial endothelial cells. Am J Physiol Lung Cell Mol Physiol 320:L726-L738 (2021). PubMed: 33565360
- Zhai W et al. Sunitinib-suppressed miR-452-5p facilitates renal cancer cell invasion and metastasis through modulating SMAD4/SMAD7 signals. Mol Cancer 17:157 (2018). PubMed: 30419914
- García-Berrocoso T et al. Single Cell Immuno-Laser Microdissection Coupled to Label-Free Proteomics to Reveal the Proteotypes of Human Brain Cells After Ischemia. Mol Cell Proteomics 17:175-189 (2018). PubMed: 29133510
- Sithole N et al. DDX17 Specifically, and Independently of DDX5, Controls Use of the HIV A4/5 Splice Acceptor Cluster and Is Essential for Efficient Replication of HIV. J Mol Biol 430:3111-3128 (2018). PubMed: 30131116
- Chen C et al. DNA-methylation-mediated repression of miR-766-3p promotes cell proliferation via targeting SF2 expression in renal cell carcinoma. Int J Cancer 141:1867-1878 (2017). PubMed: 28657135
- Roundtree IA et al. YTHDC1 mediates nuclear export of N6-methyladenosine methylated mRNAs. Elife 6:N/A (2017). PubMed: 28984244
- Rivera-Serrano EE et al. A Cytoplasmic RNA Virus Alters the Function of the Cell Splicing Protein SRSF2. J Virol 91:N/A (2017). PubMed: 28077658
- Xie N et al. SRSF1 promotes vascular smooth muscle cell proliferation through a ?133p53/EGR1/KLF5 pathway. Nat Commun 8:16016 (2017). IF ; Rat . PubMed: 28799539
- Harhouri K et al. MG132-induced progerin clearance is mediated by autophagy activation and splicing regulation. EMBO Mol Med 9:1294-1313 (2017). ICC/IF . PubMed: 28674081
- Qiao Y et al. High Glucose Stimulates Tumorigenesis in Hepatocellular Carcinoma Cells Through AGER-Dependent O-GlcNAcylation of c-Jun. Diabetes 65:619-32 (2016). PubMed: 26825459