Name: Anti-SC35 antibody [EPR12238]
Brand: RabMAb
SKU: ab204916
Rating: 4 (2 reviews)

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Immunocytochemistry/ Immunofluorescence - Anti-SC35 antibody [EPR12238] (ab204916)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR12238] to SC35
Suitable for: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WB
Reacts with: Human

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

Product name

Anti-SC35 antibody [EPR12238]
See all SC35 primary antibodies
Description

Rabbit monoclonal [EPR12238] to SC35
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WBmore details
Species reactivity

Reacts with: Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: K562, HeLa, 293 and HepG2 cell lysates; Human thymus, fetal kidney and fetal brain lysates. IHC-P: Human cervical carcinoma tissue. ICC/IF: HEK-293 and HeLa cells. IP: HeLa whole cell lysate.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR12238
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human SC35 protein (ab152674)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab204916 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
Flow Cyt (Intra)		1/400.
ICC/IF	(1)	1/100.
IP		1/30.
IHC-P		1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB	(1)	1/1000. Detects a band of approximately 35 kDa (predicted molecular weight: 25 kDa).

Notes
Flow Cyt (Intra) 1/400.
ICC/IF 1/100.
IP 1/30.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 35 kDa (predicted molecular weight: 25 kDa).

Function

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.
Sequence similarities

Belongs 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 localization

Nucleus.
Target information above from: UniProt accession Q01130 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 6427 Human
- Omim: 600813 Human
- SwissProt: Q01130 Human
- Unigene: 584801 Human
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

Immunocytochemistry/ Immunofluorescence - Anti-SC35 antibody [EPR12238] (ab204916)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293 (Human epithelial cells from embryonic kidney) cells labeling SC35 with ab204916 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HEK-293 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab204916 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Western blot - Anti-SC35 antibody [EPR12238] (ab204916)

All lanes : Anti-SC35 antibody [EPR12238] (ab204916) at 1/1000 dilution

Lane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 3 : 293 (Human epithelial cells from embryonic kidney) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 25 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?

Exposure time: 10 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Flow Cytometry (Intracellular) - Anti-SC35 antibody [EPR12238] (ab204916)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling SC35 with ab204916 at 1/400 dilution (0.1µg)/ Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SC35 antibody [EPR12238] (ab204916)

Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labeling SC35 with ab204916 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human cervical carcinoma is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-SC35 antibody [EPR12238] (ab204916)

SC35 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab204916 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab204916 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500.
Lane 1: HeLa whole cell lysate 10ug (Input). Lane 2: ab204916 IP in HeLa whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab204916 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
Western blot - Anti-SC35 antibody [EPR12238] (ab204916)

Anti-SC35 antibody [EPR12238] (ab204916) at 1/1000 dilution + HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 25 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-SC35 antibody [EPR12238] (ab204916)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling SC35 with ab204916 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab204916 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Western blot - Anti-SC35 antibody [EPR12238] (ab204916)

Anti-SC35 antibody [EPR12238] (ab204916) at 1/5000 dilution + Human thymus lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 25 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-SC35 antibody [EPR12238] (ab204916)

Anti-SC35 antibody [EPR12238] (ab204916) at 1/1000 dilution + Human fetal kidney lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 25 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-SC35 antibody [EPR12238] (ab204916)

Anti-SC35 antibody [EPR12238] (ab204916) at 1/1000 dilution + Human fetal brain lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 25 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Anti-SC35 antibody [EPR12238] (ab204916)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab204916? Please let us know so that we can cite the reference in this datasheet.

ab204916 has been referenced in 10 publications.

Tomita T et al. Extracellular mRNA transported to the nucleus exerts translation-independent function. Nat Commun 12:3655 (2021). PubMed: 34135341
Yang L et al. Mirtronic miR-4646-5p promotes gastric cancer metastasis by regulating ABHD16A and metabolite lysophosphatidylserines. Cell Death Differ 28:2708-2727 (2021). PubMed: 33875796
Chen Z et al. CircPLCE1 facilitates the malignant progression of colorectal cancer by repressing the SRSF2-dependent PLCE1 pre-RNA splicing. J Cell Mol Med 25:7244-7256 (2021). PubMed: 34173324
Francis AC et al. HIV-1 replication complexes accumulate in nuclear speckles and integrate into speckle-associated genomic domains. Nat Commun 11:3505 (2020). PubMed: 32665593
Ilik IA et al. SON and SRRM2 are essential for nuclear speckle formation. Elife 9:N/A (2020). PubMed: 33095160
Teslow EA et al. Obesity-induced MBD2_v2 expression promotes tumor-initiating triple-negative breast cancer stem cells. Mol Oncol 13:894-908 (2019). PubMed: 30636104
Gudkova D et al. HCF-2 inhibits cell proliferation and activates differentiation-gene expression programs. Nucleic Acids Res 47:5792-5808 (2019). PubMed: 31049581
Li X et al. Downregulation of SRPK2 promotes cell cycle arrest though E2F1 in non-small cell lung cancer. Eur J Histochem 63:N/A (2019). PubMed: 31833327
Chakraborty P et al. DHX9 helicase promotes R-loop formation in cells with impaired RNA splicing. Nat Commun 9:4346 (2018). PubMed: 30341290
Shakyawar DK et al. C3G dynamically associates with nuclear speckles and regulates mRNA splicing. Mol Biol Cell 29:1111-1124 (2018). PubMed: 29496966

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Application

Immunocytochemistry/ Immunofluorescence

Sample

Zebrafish Cell (Whole Embryo, Dome Stage)

Permeabilization

Yes - 100% Methanol -20°C overnight, ddH2O room temperature for 15 minutes

Specification

Whole Embryo, Dome Stage

Blocking step

(agent) for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

Fixative

Formaldehyde

Dr. Lennart Hilbert

Verified customer

Submitted Sep 23 2016

Application

Western blot

Sample

Human Cell lysate - whole cell (Human lymphocytes)

Gel Running Conditions

Reduced Denaturing (4-12% Bis-Tris)

Loading amount

40 µg

Specification

Human lymphocytes

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 27 2016

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (10)

Customer reviews and Q&As

Immunocytochemistry/ Immunofluorescence abreview for Anti-SC35 antibody [EPR12238]

Western blot abreview for Anti-SC35 antibody [EPR12238]

Post-translational
modifications