Recombinant Anti-SF3B3 antibody [EPR18441] (ab209403)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18441] to SF3B3
- Suitable for: WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-SF3B3 antibody [EPR18441]
See all SF3B3 primary antibodies -
Description
Rabbit monoclonal [EPR18441] to SF3B3 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: HeLa, Jurkat, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Human fetal liver and fetal heart lysates; Mouse brain, kidney and spleen lysates; Rat kidney and spleen lysates. IHC-P: Human colon, Human breast cancer (ER+) and Human breast cancer (ER-) tissues; Mouse cerebral cortex tissue; Rat kidney tissue. ICC/IF: C6 and RAW 264.7 cells. IP: HeLa and C6 cell lysates.
-
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
-
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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18441 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab209403 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 |
---|---|---|
WB |
1/2000. Detects a band of approximately 136 kDa (predicted molecular weight: 136 kDa).
|
|
IHC-P |
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
ICC/IF |
1/500.
|
|
IP |
1/60.
|
Notes |
---|
WB
1/2000. Detects a band of approximately 136 kDa (predicted molecular weight: 136 kDa). |
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/500. |
IP
1/60. |
Target
-
Function
Subunit of the splicing factor SF3B required for 'A' complex assembly formed by the stable binding of U2 snRNP to the branchpoint sequence (BPS) in pre-mRNA. Sequence independent binding of SF3A/SF3B complex upstream of the branch site is essential, it may anchor U2 snRNP to the pre-mRNA. May also be involved in the assembly of the 'E' complex. Belongs also to the minor U12-dependent spliceosome, which is involved in the splicing of rare class of nuclear pre-mRNA intron. -
Sequence similarities
Belongs to the RSE1 family. -
Cellular localization
Nucleus. - Information by UniProt
-
Database links
- Entrez Gene: 23450 Human
- Entrez Gene: 101943 Mouse
- Entrez Gene: 292019 Rat
- Omim: 605592 Human
- SwissProt: Q15393 Human
- SwissProt: Q921M3 Mouse
- Unigene: 514435 Human
- Unigene: 236123 Mouse
-
Alternative names
- KIAA0017 antibody
- Pre mRNA splicing factor SF3b 130 kDa subunit antibody
- Pre-mRNA-splicing factor SF3b 130 kDa subunit antibody
see all
Images
-
All lanes : Anti-SF3B3 antibody [EPR18441] (ab209403) at 1/5000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 136 kDa
Observed band size: 136 kDa
Exposure time: 8 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
-
Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling SF3B3 with ab209403 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nucleus staining on rat kidney 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.
-
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (Rat glial tumor cell line) cells labeling SF3B3 with ab209403 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on C6 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab209403 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
-
SF3B3 was immunoprecipitated from 1mg / 300µl of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab209403 at 1/60 dilution (5µg/ 1mg of lysate).
Western blot was performed from the immunoprecipitate using ab209403 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa whole cell lysate, 10ug (Input).
Lane 2: ab209403 IP in HeLa whole cell lysate.
Lane 3: Rabbit IgG,monoclonal - Isotype Control (ab172730) instead of ab209403 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
-
All lanes : Anti-SF3B3 antibody [EPR18441] (ab209403) at 1/2000 dilution
Lane 1 : Human fetal liver lysate
Lane 2 : Human fetal heart lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 136 kDa
Observed band size: 136 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
-
All lanes : Anti-SF3B3 antibody [EPR18441] (ab209403) at 1/2000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Rat kidney lysate
Lane 5 : Rat spleen lysate
Lane 6 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 7 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 8 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 9 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 136 kDa
Observed band size: 136 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time : Lane 1/2/4: 3 minutes; Lane 3/5: 30 seconds; Lane 6/7/8/9: 8 seconds.
-
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling SF3B3 with ab209403 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nucleus staining on Human colon 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.
-
Immunohistochemical analysis of paraffin-embedded Human breast cancer (ER+) tissue labeling SF3B3 with ab209403 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nucleus staining on ER+ breast cancer 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.
-
Immunohistochemical analysis of paraffin-embedded Human breast cancer (ER-) tissue labeling SF3B3 with ab209403 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Weak nucleus staining on ER- breast cancer 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.
-
Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling SF3B3 with ab209403 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nucleus staining on mouse cerebral cortex 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.
-
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling SF3B3 with ab209403 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on RAW 264.7 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody- Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG(AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab209403 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
-
SF3B3 was immunoprecipitated from 1mg / 300µl of C6 (Rat glial tumor cell line) whole cell lysate with ab209403 at 1/60 dilution (5µg / 1mg of lysate).
Western blot was performed from the immunoprecipitate using ab209403 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: C6 whole cell lysate, 10ug (Input).
Lane 2: ab209403 IP in C6 whole cell lysate.
Lane 3: Rabbit IgG,monoclonal - Isotype Control (ab172730) instead of ab209403 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (1)
ab209403 has been referenced in 1 publication.
- Cretu C et al. Structural Basis of Splicing Modulation by Antitumor Macrolide Compounds. Mol Cell 70:265-273.e8 (2018). PubMed: 29656923