Overview

  • Product name
    Anti-SSB antibody [mAbcam75927]
    See all SSB primary antibodies
  • Description
    Mouse monoclonal [mAbcam75927] to SSB
  • Tested applications
    Suitable for: ICC/IF, IP, Flow Cyt, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human SSB aa 300 to the C-terminus (C terminal).
    (Peptide available as ab106476)

  • Positive control
    • This antibody gave a positive signal in the following lysates: HEK293 Whole Cell; HeLa Nuclear; Raji Nuclear; MCF7 Nuclear, MCF7 Whole Cell; Ramos Whole Cell; Human Fetal Heart Tissue. This antibody gave a positive result in IHC in the following FFPE tissue: Human breast adenocarcinoma.
  • General notes

    This antibody clone is manufactured by Abcam.

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.
  • Storage buffer
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Please note that some batches of ab75927 may contain 0.4M arginine. Please contact Scientific Support for further information.
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Monoclonal
  • Clone number
    mAbcam75927
  • Myeloma
    Sp2/0-Ag14
  • Isotype
    IgG2a
  • Research areas

Applications

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 Use a concentration of 5 µg/ml.
IP Use a concentration of 5 µg/ml.
Flow Cyt Use 0.5µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
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.

Target

  • 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
    modifications
    Phosphorylated. The phosphorylation sites are at the C-terminal part of the protein.
    The N-terminus is blocked.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Autoantigen La antibody
    • La antibody
    • La autoantigen antibody
    • La autoantigen homolog antibody
    • La protein antibody
    • La ribonucleoprotein antibody
    • La ribonucleoprotein domain family member 3 antibody
    • LA_HUMAN antibody
    • LARP3 antibody
    • Lupus La antigen antibody
    • Lupus La protein antibody
    • Lupus La protein homolog antibody
    • MGC118101 antibody
    • MGC93380 antibody
    • mRNA for autoantigen antibody
    • OTTMUSP00000014043 antibody
    • RP23 273G23.1 antibody
    • Sjoegren syndrome type B antigen antibody
    • Sjogren syndrome antigen B (autoantigen La) antibody
    • Sjogren syndrome antigen B antibody
    • SS B antibody
    • SS-B antibody
    • SS-B/La protein antibody
    • SSB antibody
    see all

Anti-SSB antibody [mAbcam75927] images

  • 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

  • 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

  • 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
    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
  • ICC/IF image of ab75927 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 ab75927 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse IgG (H+L)(ab96879) 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. This antibody also gave a positive result in Hek293, HepG2, and MCF-7 cell types Methanol fixed (100%, 5 min) at 5ug/ml


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 47 kDa
  • 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.
  • 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.

References for Anti-SSB antibody [mAbcam75927] (ab75927)

This product has been referenced in:
  • 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). Read more (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 . Read more (PubMed: 27446609) »

See all 4 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293 cells)
Gel Running Conditions
Reduced Denaturing (12,5 %)
Loading amount
20 µg
Specification
HEK293 cells
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Dr. Melanie Thelen

Verified customer

Submitted May 23 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (PBMC)
Loading amount
30 µg
Specification
PBMC
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted May 14 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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