Anti-SC35 antibody [SC-35] ab11826 is a mouse monoclonal antibody that is used in SC35 immunofluorescence. Suitable for human, mouse and rat samples.
- Antibody clone SC-35 is the most widely used clone for SC35 on the market and is cited in >300 publications
IgG1
Mouse
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 93% PBS, 6.97% L-Arginine
Liquid
Monoclonal
WB | ICC/IF | |
---|---|---|
Human | Not recommended | Tested |
Mouse | Not recommended | Tested |
Rat | Not recommended | Tested |
Drosophila melanogaster | Not recommended | Predicted |
Newt | Not recommended | Predicted |
Rhesus monkey | Not recommended | Predicted |
Xenopus laevis | Not recommended | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes Lysis buffer: 6.0% (w/v) SDS, 0.14 mol Tris (pH 6.8), 22.4% (v/v) glycerol, protease inhibitor cocktail. Centrifuge at 14 000 g for 15 min at 48C. Block non-specific binding sites with 5% (w/v) skimmed milk powder in TBS with 0.1% (v/v) Tween 20 for 1 h. Incubate blots with primary antibody (diluted in 5% (w/v) skimmed milk powder in TBS with 0.1% (v/v) Tween 20) for 1 h at room temperature. |
Species Rat | Dilution info - | Notes Lysis buffer: 6.0% (w/v) SDS, 0.14 mol Tris (pH 6.8), 22.4% (v/v) glycerol, protease inhibitor cocktail. Centrifuge at 14 000 g for 15 min at 48C. Block non-specific binding sites with 5% (w/v) skimmed milk powder in TBS with 0.1% (v/v) Tween 20 for 1 h. Incubate blots with primary antibody (diluted in 5% (w/v) skimmed milk powder in TBS with 0.1% (v/v) Tween 20) for 1 h at room temperature. |
Species Human | Dilution info - | Notes Lysis buffer: 6.0% (w/v) SDS, 0.14 mol Tris (pH 6.8), 22.4% (v/v) glycerol, protease inhibitor cocktail. Centrifuge at 14 000 g for 15 min at 48C. Block non-specific binding sites with 5% (w/v) skimmed milk powder in TBS with 0.1% (v/v) Tween 20 for 1 h. Incubate blots with primary antibody (diluted in 5% (w/v) skimmed milk powder in TBS with 0.1% (v/v) Tween 20) for 1 h at room temperature. |
Species Xenopus laevis | Dilution info - | Notes - |
Species Drosophila melanogaster | Dilution info - | Notes - |
Species Rhesus monkey | Dilution info - | Notes - |
Species Newt | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 5 µg/mL | Notes - |
Species Rat | Dilution info 5 µg/mL | Notes - |
Species Human | Dilution info 5 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Xenopus laevis, Drosophila melanogaster, Rhesus monkey, Newt | Dilution info - | Notes - |
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Necessary for the splicing of pre-mRNA. It is required for formation of the earliest ATP-dependent splicing complex and interacts with spliceosomal components bound to both the 5'- and 3'-splice sites during spliceosome assembly. It also is required for ATP-dependent interactions of both U1 and U2 snRNPs with pre-mRNA. 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. Binds to purine-rich RNA sequences, either 5'-AGSAGAGTA-3' (S=C or G) or 5'-GTTCGAGTA-3'. Can bind to beta-globin mRNA and commit it to the splicing pathway. The phosphorylated form (by SRPK2) is required for cellular apoptosis in response to cisplatin treatment.
Serine/arginine-rich splicing factor 2, Protein PR264, Splicing factor SC35, SC-35, SRSF2, SFRS2
Anti-SC35 antibody [SC-35] ab11826 is a mouse monoclonal antibody that is used in SC35 immunofluorescence. Suitable for human, mouse and rat samples.
- Antibody clone SC-35 is the most widely used clone for SC35 on the market and is cited in >300 publications
IgG1
Mouse
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 93% PBS, 6.97% L-Arginine
Liquid
Monoclonal
SC-35
Affinity purification Protein G
This antibody recognizes a phospho-epitope of the non-snRNP (small nuclear ribonucleoprotein particles) factor SC35. The antibody reacts with the splicing factor SC-35 and with the SC-35-related non-snRNP factor SF2/ASF. Recent data suggests this clone may cross-react with additional proteins within the spliceosome complex (PMID: 33095160)
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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This supplementary information is collated from multiple sources and compiled automatically.
SC35 also known as SRSF2 (serine/arginine-rich splicing factor 2) is an essential splicing factor with a molecular mass around 26 kDa. This protein is part of the SR protein family which are known for their roles in splicing regulation due to their serine/arginine-rich domains. SC35 predominantly localizes in the nucleus of cells where it accumulates in nuclear speckles areas rich in splicing factors. Biologists often use immunofluorescence techniques and anti-SC antibodies to visualize and study SC35's localization and function within cells.
SC35 influences various aspects of mRNA processing particularly pre-mRNA splicing. The protein functions as a splicing enhancer by interacting with pre-mRNA at specific sites to assist exon inclusion during splicing. SC35 acts as part of the spliceosomal complex which is critical during the assembly and rearrangement of the spliceosome. Additionally it interacts with other splicing factors to ensure accurate and efficient splicing which is fundamental for gene expression and consequent protein synthesis.
The SC35 protein participates in the RNA splicing pathway a significant process in mRNA maturation and gene expression regulation. This protein interacts with other SR proteins like SRSF1 and components of the spliceosomal machinery. SC35's function is also important in the alternative splicing pathway impacting the diversity of mRNA transcripts through the production of different exon combinations. By regulating alternative splicing SC35 helps produce protein diversity vital for numerous cellular processes.
Changes in SC35 expression or function have been linked to several human conditions. Notably its dysregulation can contribute to cancer development through altered splicing patterns that impact oncogene and tumor suppressor gene expression. Furthermore SC35 abnormalities are associated with spinal muscular atrophy a neurodegenerative disorder where splicing regulation plays a significant role. In these diseases SC35 may interact with other proteins such as SMN (survival of motor neuron protein) and impact their functions highlighting its relevance in maintaining cellular health.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Immunocytochemistry/ Immunofluorescence analysis of HEK-293T and RKO cells transiently transfected with CDX2/AS-His and co-stained for CDX2/AS-His and SC35 (ab11826). All proteins localized to the nucleus and merged images revealed co-localization of CDX2/AS with SC35.
Immunocytochemistry/ Immunofluorescence analysis of untransfected U-2 OS cells (A) and cells transfected with HSV US1 or US1.5 fixed and stained for FLAG (red) and SC35 (green) to identify viral proteins and nuclear speckles respectively. Transfected cells were fixed 40 h post transfection with 3.7% formaldehyde in PBS (20 min), permeabilized with 0.5% Triton X-100 in PBS (10 min), and blocked with 4% BSA in PBS (20 min) prior to incubation with Anti-SC35 antibody [SC-35] - Nuclear Speckle Marker (ab11826) and secondary antibodies in 4% BSA in PBS. DAPI was used for visualization of nuclear DNA. Scale bar = 10 μm.
Immunocytochemistry/ Immunofluorescence analysis of human adenocarcinoma HT-29 (Human colorectal adenocarcinoma cell line) cells labeling SC35 with ab11826 at 1/200 dilution. Cells were fixed in paraformaldehyde and permeabilized with Triton X-100 and Saponin. Blocking of the cells was done with 1% BSA for 1 hour at 37°C; staining with ab11826 at 1/200 was carried out for 16 hours at 4°C in PBS buffer. An anti-mouse IgG3 (Alexa Fluor® 594) secondary antibody was used at 1/200 dilution.
ab11826 staining SC35 in human fibroblast cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.3% Triton X-100 in PBS and blocked with 5% Normal Goat Serum/0.3% Triton X-100 in PBS for 60 minutes at 25°C. Samples were incubated with primary antibody (1/500 in 1% BSA/ 0.3% Triton X-100 in PBS) for 16 hours at 4°C. An Alexa Flour® 488 goat anti-mouse IgG (H+L), F(ab')2 Fragment Ig was used as the secondary antibody at a dilution of 1/1000.
HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were fixed 24–48 hours after transfection using 4% paraformaldehyde, permeabilized with 0.2% triton X-100/PBS and probed with ab11826 followed by FITC conjugated secondary antibodies (green). After washing with PBS, slides were mounted using Citifluor and analysed by confocal microscopy. Cells were visualized under a Leica laser scanning confocal microscope equipped with a DM-RXE microscope and an argon-krypton laser.
ab11826 staining SC35 - Nuclear Speckle Marker in Rin-5F cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab11826 at 5μg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
ab11826 staining SC35 - Nuclear Speckle Marker in NIH3T3 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab11826 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
ab11826 staining SC35 - Nuclear Speckled Marker in MCF7 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab11826 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Immunocytochemistry/ Immunofluorescence analysis of HEK-293 human kidney cells labeling SC35 with ab11826 at 1/400 dilution. Cells were fixed with methanol and blocked with PBS for 1 hour at 4°C. Staining with ab11826 was carried out in PBS buffer for 2 hours at 4°C. An undiluted goat anti-mouse Alexa Fluor® 594 secondary antibody was used.
Immunocytochemistry/ Immunofluorescence analysis of human hippocampus cells labeling SC35 with ab11826 at 1/200 dilution. Cells were fixed with formaldehyde and blocked with PBS for 1 hour at 4°C. Staining with ab11826 was carried out in PBS buffer for 12 hours at 4°C. A goat anti-mouse Alexa Fluor® 594 secondary antibody was used at 1/1000 dilution.
ab11826 (1/1000) staining SC35 (phospho) in human retinal pigment epithelial (RPE) cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (blue). Please refer to abreview for further experimental details.
ab11826 staining cultured human colon adenocarcinoma HT-29 cells.
Cells were PFA fixed and permeabilized in Triton X-100 and saponin prior to blocking with 1% BSA for 1 hour at RT. The primary antibody was diluted 1/200 and incubated with the sample for 16 hours at 4°C. An Alexa Fluor® 594 conjugated goat anti-mouse IgG3 antibody was used as the secondary.
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