Anti-SSB antibody [mAbcam75927] (ab75927)
Key features and details
- Mouse monoclonal [mAbcam75927] to SSB
- Suitable for: ICC/IF, IP, WB, IHC-P, Flow Cyt (Intra)
- Reacts with: Human
- Isotype: IgG2a
Overview
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Product name
Anti-SSB antibody [mAbcam75927]
See all SSB primary antibodies -
Description
Mouse monoclonal [mAbcam75927] to SSB -
Host species
Mouse -
Tested applications
Suitable for: ICC/IF, IP, WB, IHC-P, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide corresponding to Human SSB aa 300 to the C-terminus (C terminal).
(Peptide available asab106476) -
Positive control
- WB: HEK293 Whole Cell; HeLa Nuclear; Raji Nuclear; MCF7 Nuclear, MCF7 Whole Cell; Ramos Whole Cell; Human Fetal Heart Tissue. IHC: Human breast adenocarcinoma. ICC: HeLa cells
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 long term. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
mAbcam75927 -
Myeloma
Sp2/0-Ag14 -
Isotype
IgG2a -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab75927 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/IF | (1) |
Use a concentration of 5 µg/ml.
|
IP |
Use a concentration of 5 µg/ml.
|
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WB | (3) |
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
|
IHC-P |
Use a concentration of 5 µg/ml.
|
|
Flow Cyt (Intra) |
Use 0.5µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
Notes |
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ICC/IF
Use a concentration of 5 µg/ml. |
IP
Use a concentration of 5 µg/ml. |
WB
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa). |
IHC-P
Use a concentration of 5 µg/ml. |
Flow Cyt (Intra)
Use 0.5µg for 106 cells. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
Target
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Function
Binds to the 3' poly(U) terminii of nascent RNA polymerase III transcripts, protecting them from exonuclease digestion and facilitating their folding and maturation. -
Sequence similarities
Contains 1 HTH La-type RNA-binding domain.
Contains 1 RRM (RNA recognition motif) domain. -
Post-translational
modificationsPhosphorylated. The phosphorylation sites are at the C-terminal part of the protein.
The N-terminus is blocked. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 6741 Human
- GenBank: NP_003133.1 Human
- Omim: 109090 Human
- SwissProt: P05455 Human
- Unigene: 632535 Human
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Alternative names
- Autoantigen La antibody
- La antibody
- La autoantigen antibody
see all
Images
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All lanes : Anti-SSB antibody [mAbcam75927] (ab75927) at 5 µg/ml
Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lane 3 : Raji (Human Burkitt's lymphoma cell line) Nuclear Lysate - tumor cell line (ab30127)
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) Nuclear Lysate
Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 6 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate
Lane 7 : Heart (Human) Whole Cell Lysate - fetal normal tissue
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 47 kDa
Observed band size: 47 kDa
Exposure time: 30 seconds -
IHC image of SSB staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab75927, 0.5µg/ml overnight at +4°C. An HRP-conjugated secondary (Ab97245, 1/200 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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SSB was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Mouse monoclonal to SSB and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab75927.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.
Band: 47kDa; SSB
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ab75927 staining SSB in HeLa 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 ab75927 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and 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 100% methanol (5 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Anti-SSB antibody [mAbcam75927] (ab75927) at 1 µg/ml +
Recombinant Human SSB protein (ab84477) at 0.1 µg
Secondary
Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 47 kDa
Exposure time: 30 seconds -
Overlay histogram showing Ramos cells stained with ab75927 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75927, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Ramos cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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IHC image of SSB staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab75927, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab75927 has been referenced in 10 publications.
- Huang X et al. La protein regulates protein expression by binding with the mRNAs of target genes and participates the pathological process of ovarian cancer. Front Oncol 12:763480 (2022). PubMed: 36110943
- Butler CL et al. Discovery of non-HLA antibodies associated with cardiac allograft rejection and development and validation of a non-HLA antigen multiplex panel: From bench to bedside. Am J Transplant 20:2768-2780 (2020). PubMed: 32185871
- Pollak AJ et al. Gapmer Antisense Oligonucleotides Targeting 5S Ribosomal RNA Can Reduce Mature 5S Ribosomal RNA by Two Mechanisms. Nucleic Acid Ther 30:312-324 (2020). PubMed: 32589504
- Mandal A & Chaubey B Expression of Duplex shRNAs through a Lentiviral Vector against Cellular and Viral Genes Inflicts Sustained Inhibition of Hepatitis C Virus Replication. Intervirology 61:79-91 (2018). PubMed: 30253401
- Liang XH et al. Translation can affect the antisense activity of RNase H1-dependent oligonucleotides targeting mRNAs. Nucleic Acids Res 46:293-313 (2018). PubMed: 29165591
- Liang XH et al. RNase H1-Dependent Antisense Oligonucleotides Are Robustly Active in Directing RNA Cleavage in Both the Cytoplasm and the Nucleus. Mol Ther 25:2075-2092 (2017). PubMed: 28663102
- Gao W et al. La Autoantigen Induces Ribosome Binding Protein 1 (RRBP1) Expression through Internal Ribosome Entry Site (IRES)-Mediated Translation during Cellular Stress Condition. Int J Mol Sci 17:N/A (2016). PubMed: 27447629
- Mandal A et al. Combinations of siRNAs against La Autoantigen with NS5B or hVAP-A Have Additive Effect on Inhibition of HCV Replication. Hepat Res Treat 2016:9671031 (2016). WB . PubMed: 27446609
- Shwetha S et al. HuR Displaces Polypyrimidine Tract Binding Protein To Facilitate La Binding to the 3' Untranslated Region and Enhances Hepatitis C Virus Replication. J Virol 89:11356-71 (2015). WB ; Human . PubMed: 26339049
- Spurlock CF et al. Defective structural RNA processing in relapsing-remitting multiple sclerosis. Genome Biol 16:58 (2015). WB . PubMed: 25885816