Anti-SC35 antibody [SC-35] - Nuclear Speckle Marker (ab11826)

Overview

  • Product nameAnti-SC35 antibody [SC-35] - Nuclear Speckle Marker
    See all SC35 primary antibodies
  • Description
    Mouse monoclonal [SC-35] to SC35 - Nuclear Speckle Marker
  • SpecificityIn immunohistology and immunoelectronmicroscopy, the antibody labels SC-35 as a speckled pattern that occupies a portion of the nucleoplasm, excluding the nucleoli. Nie et al (2002) report that "In human endometrium the SC35 protein was detected by western blot analysis and had a molecular mass of approximately 90 kDa, indicating that in the endometrial extracts, the 35 kDa protein remains bound to one or more components of the spliceosome."
  • Tested applicationsSuitable for: ICC, ICC/IF, Other, IHC-P, ELISA, IP, IHC-Frmore details
    Unsuitable for: WB
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Xenopus laevis, Drosophila melanogaster, Rhesus monkey, Newt
  • EpitopeThis 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.
  • Positive control
    • Cultured human fibroblasts. This antibody gave a positive signal in the following cell types: HeLa. (IF)

Properties

Applications

Our Abpromise guarantee covers the use of ab11826 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
ICC 1/2000.
ICC/IF Use a concentration of 1 µg/ml.
Other Use at an assay dependent concentration.
IHC-P 1/500.
ELISA Use at an assay dependent concentration.
AP Use at an assay dependent concentration.
IP Use at an assay dependent concentration. See Fu and Maniatis 1992 reference.
IHC-Fr Use at an assay dependent concentration. PubMed: 24098751
  • Application notesIs unsuitable for WB.
  • Target

    • FunctionNecessary 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.
    • Sequence similaritiesBelongs to the splicing factor SR family.
      Contains 1 RRM (RNA recognition motif) domain.
    • Post-translational
      modifications
      Extensively phosphorylated on serine residues in the RS domain.
    • Cellular localizationNucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • 35 kDa antibody
      • arginine/serine-rich 2 antibody
      • PR264 antibody
      • Protein PR264 antibody
      • SC 35 antibody
      • SC-35 antibody
      • SC35 antibody
      • Serine/arginine-rich splicing factor 2 antibody
      • SFRS 2 antibody
      • SFRS2 antibody
      • SFRS2A antibody
      • Splicing component 35 kDa antibody
      • Splicing component antibody
      • Splicing factor antibody
      • Splicing factor arginine/serine rich 2 antibody
      • Splicing factor SC35 antibody
      • Splicing speckle antibody
      • Splicing speckles antibody
      • SR splicing factor 2 antibody
      • SRp30b antibody
      • SRSF2 antibody
      • SRSF2_HUMAN antibody
      see all

    Anti-SC35 antibody [SC-35] - Nuclear Speckle Marker images

    • Immunocytochemistry/ Immunofluorescence analysis of 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hippocampus tissue sections labeling SC35 with ab11826 at 1/500 dilution. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with a pH 9 buffer. Blocking of the sample was done with 100%PBS for 1 hour at 25°C, followed by staining with ab11826 at 1/500 in PBS for 14h at 4°C. A biotinylated goat anti-mouse polyclonal antibody at 1/200 was used as the secondary antibody.  

      See Abreview

    • Immunocytochemistry/ Immunofluorescence analysis of HEK293 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. 

      See Abreview

    • Immunocytochemistry/ Immunofluorescence analysis of untransfected U2OS 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 HT29 cells labeling SC35 with ab11826 at 1/200 dilution. Cells were fixed in paraformaldehyde and permeabilized with Triton X100 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. 

      See Abreview

    • 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. 

      See Abreview

    • Immunohistochemistry (Frozen Sections) analysis of human retinal sections (16 µm cryosections) labelling SC35 with ab11826 at 1/300 dilution (red). The cryosections were first hydrated in phosphate-buffered saline (PBS, pH 7.4) and washed three times (5 minutes each at room temperature (RT)), followed by incubation with PBTS buffer (1X PBS with 0.1% triton-X 100, 0.2% BSA and 0.02% SDS) for an hour at RT. Primary antibody Anti-SC35 antibody [SC-35] - Nuclear Speckle Marker (ab11826) was incubated in PBTS buffer overnight at 4°C. Sections were washed with PBTS buffer containing DAPI 10 times (15 minutes each at RT). Following the washes, secondary antibody anti-mouse antibody conjugated with Alexa Fluor® at 1/750 dilution was incubated in PBTS buffer overnight at 4°C. Sections were washed with PBTS buffer 7 times (15 minutes each at RT), rinsed with PBS and covered with anti-fade reagent and coverslip glass.

    • 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.

      See Abreview

    • ab11826 (1/1000) staining SC35 (phospho) in Human retinal pigment epithelial (RPE) cells (green). Cells were fixed in paraformaldehyde, permeabilised 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.

      See Abreview

    • ab11826 staining cultured human colon adenocarcinoma HT29 cells.  Cells were PFA fixed and permeabillized in Triton X100 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.

      See Abreview

    • HeLa cells were fixed 24–48 hours after transfection using 4% paraformaldehyde, permeabilised 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 visualised under a Leica laser scanning confocal microscope equipped with a DM-RXE microscope and an argon-krypton laser.
    • ICC/IF image of ab11826 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab11826 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    References for Anti-SC35 antibody [SC-35] - Nuclear Speckle Marker (ab11826)

    This product has been referenced in:
    • Deane CAS & Brown IR Differential Targeting of Hsp70 Heat Shock Proteins HSPA6 and HSPA1A with Components of a Protein Disaggregation/Refolding Machine in Differentiated Human Neuronal Cells following Thermal Stress. Front Neurosci 11:227 (2017). ICC/IF ; Human . Read more (PubMed: 28484369) »
    • Taylor SE  et al. Differential sub-nuclear distribution of hypoxia-inducible factors (HIF)-1 and -2 alpha impacts on their stability and mobility. Open Biol 6:N/A (2016). ICC/IF ; Human . Read more (PubMed: 27655733) »

    See all 39 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Human Tissue sections (Hippocampus-Paraffin embedded)
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: pH9
    Permeabilization No
    Specification Hippocampus-Paraffin embedded
    Blocking step Scy-tek Superblock as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jul 16 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (fibroblast)
    Permeabilization Yes - block contains 0.3% tritonX-100 (1H, RT)
    Specification fibroblast
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jun 29 2016

    Application Western blot
    Sample Human Cell lysate - whole cell (HEK293 (kidney cell))
    Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris Gradient)
    Loading amount 5 µg
    Specification HEK293 (kidney cell)
    Blocking step Licor Blocking Reagent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C
    Username

    Abcam user community

    Verified customer

    Submitted Jul 18 2015

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step Scytek Superblock as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C
    Sample Human Cell (HEK293 (kidney))
    Specification HEK293 (kidney)
    Permeabilization No
    Fixative Methanol
    Username

    Dr. Sam Nowitzki

    Verified customer

    Submitted Apr 06 2015

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step Scytek Superblock as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C
    Antigen retrieval step Heat mediated
    Sample Human Tissue sections (Hippocampus (Brain))
    Specification Hippocampus (Brain)
    Permeabilization No
    Fixative Formaldehyde
    Username

    Dr. Sam Nowitzki

    Verified customer

    Submitted Mar 31 2015

    After contacting the laboratory, the IHC-P protocol in the paper through the link below was used successfully with this antibody.

    http://www.reproduction-online.org/content/124/2/209.full.pdf

    Our recommended ISH protocol is given thro...

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    Thank you for contacting us again.

    we will gladly provide a free of charge replacement, however, we need to know which antibody we shall send to you.

    The unwanted cross-reactivity is present in all lots, therefore, we cannot provid...

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    Thank you for your nice reply.

    Unfortunately, it is not only a batch problem, but will happen all the time with this antibody. That's why we removed the application and references for WB. this means unfortunately, that I cannot send you a rep...

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    This antibody works fine for ICC, but we found that in WB it shows a possible cross-reactivity with the SC35-related non-snRNP factor SF2/ASF (p75) and Ad2 proteins. This may explain your unexpected results.

    Perdona por la espera.

    La mejor manera de evitar bandas inespecíficas es asegurar unas condiciones de preparación de la muestra idóneas para el anticuerpo en cuestión. Este anticuerpo reconoce el epítopo en ...

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