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Our Abpromise guarantee covers the use of ab31013 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use at an assay dependent concentration.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
|IP||Use at 1 µg/mg of lysate.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 56 kDa.|
|IHC-P||1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Staining requires boiling tissue sections in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min.|
|IHC-Fr||Use at an assay dependent concentration. PubMed: 20223936|
ab31013 staining TGF beta Receptor I in Human intestine tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% BSA for 1 hour at room temperature; antigen retrieval was by heat mediation using Abcam Heat mediated solution ab973. Samples were incubated with primary antibody (1/100) for 20 hours at 4°C. A HRP-conjugated Goat polyclonal (1/500) was used as the secondary antibody.
ab31013 staining the TGF beta Receptor in Mouse liver immortalized cells by Flow Cytometry. Cells were cultured in 5% mouse serum + PBS. The sample was incubated with the primary antibody (1/10 in PBS + 5% mouse serum) for 40 minutes at 4°C. A Alexa Fluor 594-conjugated Mouse anti-rabbit (1/1000) was used as the secondary antibody.
Gating Strategy: Negative control without antibody
ab31013 staining TGF beta Receptor I in Human Lymph node tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 20°C. Samples were incubated with primary antibody (1/50) for 12 hours at 4°C. A HRP-conjugated Goat anti-rabbit polyclonal (1/200) was used as the secondary antibody.
This image is courtesy of an anonymous AbreviewBlocking Step: 5% Milk for 1 hour at 4°C
This image is courtesy of an anonymous AbreviewBlockng Step: 5% Milk for 1 hour at 20°C
ab31013 at 1/50 staining rat heart tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step was performed in citrate buffer. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
Intense staining of rat heart valve was seen in fresh cut sections compared to weaker staining in those that had been cut 2 weeks previously. Primate placenta was used as a positive control as Abcam recommends - we saw a similar staining pattern to image already on their product page (i.e. clear positive staining was confirmed in the placental outer trophoblast layers). Negative controls (substitution of primary for buffer and suitable isotype) were clear.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"