Product nameAnti-Thrombospondin 1 antibody
DescriptionRabbit polyclonal to Thrombospondin 1
Tested applicationsSuitable for: IHC-P, ICC/IF, IP, WBmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat, Chicken, Cow
- This antibody gave a positive signal in HUVEC whole cell lysate. IHC-P: FFPE human breast adenocarcinoma tissue sections. ICC/IF: HeLa cells.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab85762 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||Use a concentration of 5 µg/ml.|
|IP||Use at an assay dependent concentration.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 155 kDa (predicted molecular weight: 129 kDa). Human Thrombospondin-1 is predicted to migrate in WB to ~155kDa as it contains a number of potential glycosylation sites; Swiss Prot predicts a protein of 129kDa not taking modifications into account.|
RelevanceThrombospondin is a regulator of many biological processes including cell growth, adhesion, migration, platelet aggregation, and fibrin deposition and lysis. It interacts with a number of plasma proteins such as fibrinogen and plasminogen and co-polymerizes with fibrin in clot formation. Thrombospondin also has multiple binding sites that interact with molecules such as fibronectin, collagens, laminin and heparan sulphate proteoglycans as well as binding growth factors such as TGF-beta1. Thrombospondin exerts an anti-adhesive effect which leads to cell rounding and detachment.
- THBS1 antibody
- TSP antibody
- Tsp1 antibody
Anti-Thrombospondin 1 antibody (ab85762) at 1 µg/ml + HUVEC (Human Umbilical Vein Endothelial Cell) Whole Cell Lysate at 10 µg
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 129 kDa
Observed band size: 155 kDa why is the actual band size different from the predicted?
Exposure time: 5 minutes
Human Thrombospondin-1 is predicted to migrate in WB to ~155kDa as it contains a number of potential glycosylation sites; Swiss Prot predicts a protein of 129kDa not taking modifications into account.
IHC image of Thrombospondin staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond 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 ab85762, 1µg/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.
ab85762 staining Thrombospondin 1 in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% 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 with the antibody ab85762 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.
ab85762 used in Immunoprecipitation.
Whole cell lysate prepared from human endothelial cells loaded at 600µg total protein.Immunoprecipitation step performed using Protein A.
For IP: ab85762 used at 3.33µg/mg lysate.For WB: ab85762 used at a1/1000 dilution.
This product has been referenced in:
- Smeda M et al. Dual antiplatelet therapy with clopidogrel and aspirin increases mortality in 4T1 metastatic breast cancer-bearing mice by inducing vascular mimicry in primary tumour. Oncotarget 9:17810-17824 (2018). Read more (PubMed: 29707148) »
- Anene C et al. Platelet microparticle delivered microRNA-Let-7a promotes the angiogenic switch. Biochim Biophys Acta 1864:2633-2643 (2018). WB . Read more (PubMed: 29684582) »