Journal: F1000Research

Article Title: A guide to selecting high-performing antibodies for STXBP1 (UniProt ID: P61764 ) for use in western blot, immunoprecipitation, and immunofluorescence

doi: 10.12688/f1000research.160174.1

Figure Lengend Snippet: Lysates of U-87 MG WT and STXBP1 KO were prepared, and 40 μg of protein were processed for western blot with the indicated STXBP1 antibodies. The Ponceau stained transfers of each blot are presented to show equal loading of WT and KO lysates and protein transfer efficiency from the acrylamide gels to the nitrocellulose membrane. Antibody dilutions were chosen according to the recommendations of the antibody supplier. Antibody dilution used: ab109023** at 1/1000, ab124920** at 1/10 000, ab315893** at 1/1000, A5420 at 1/1000, NBP3-15097** at 1/2000, AF5675 at 1/200, 13414** at 1/1000, GTX114808 at 1/500, GTX114809 at 1/500, 67137-1-Ig* at 1/5000, MA5-35796** at 1/2000, PA1-742 at 1/500 (2 μg/ml). Predicted band size: 67.5 kDa. **Recombinant antibody, *Monoclonal antibody.

Article Snippet: Bio-Techne (Novus Biologicals) , NBP3-15097 , 230461 , AB_3094817 , recombinant mono , S05-4B6 , rabbit , n/a , Wb, IF.

Techniques: Western Blot, Staining, Membrane, Recombinant

Journal: F1000Research

Article Title: A guide to selecting high-performing antibodies for STXBP1 (UniProt ID: P61764 ) for use in western blot, immunoprecipitation, and immunofluorescence

doi: 10.12688/f1000research.160174.1

Figure Lengend Snippet: U-87 MG lysates were prepared, and immunoprecipitation was performed using 0.6 mg of lysate and 2.0 μg of the indicated STXBP1 antibodies pre-coupled to Dynabeads protein A or protein G. Samples were washed and processed for western blot with the indicated STXBP1 antibody. For western blot, NBP3-15097** was used at 1/2000. The Ponceau stained transfers of each blot are shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate, HC= antibody heavy chain. **Recombinant antibody, *Monoclonal antibody.

Article Snippet: Bio-Techne (Novus Biologicals) , NBP3-15097 , 230461 , AB_3094817 , recombinant mono , S05-4B6 , rabbit , n/a , Wb, IF.

Techniques: Immunoprecipitation, Western Blot, Staining, Recombinant

Journal: F1000Research

Article Title: A guide to selecting high-performing antibodies for STXBP1 (UniProt ID: P61764 ) for use in western blot, immunoprecipitation, and immunofluorescence

doi: 10.12688/f1000research.160174.1

Figure Lengend Snippet: U-87 MG WT and STXBP1 KO cells were labelled with a green or a far-red fluorescent dye, respectively. WT and KO cells were mixed and plated to a 1:1 ratio on coverslips. Cells were stained with the indicatedSTXBP1 antibodies and with the corresponding Alexa-fluor 555 coupled secondary antibody including DAPI. Acquisition of the blue (nucleus-DAPI), green (WT), red (antibody staining) and far-red (KO) channels was performed. Representative images of the merged blue and red (grayscale) channels are shown. WT and KO cells are outlined with green and magenta dashed line, respectively. When an antibody was recommended for immunofluorescence by the supplier, we tested it at the recommended dilution. The rest of the antibodies were tested at 1 and 2 μg/ml and the final concentration was selected based on the detection range of the microscope used and a quantitative analysis not shown here. Antibody dilution used: ab109023** at 1/100, ab124920** at 1/100, ab315893** at 1/500, A5420 at 1/50, AF5675 at 1/200, NBP3-15097** at 1/500, 13414** at 1/100, GTX114808 at 1/800, GTX114809 at 1/1000, 67137-1-Ig* at 1/2000, MA5-35796** at 1/100, PA1-742 at 1/1000. Bars = 10 μm. **Recombinant antibody, *Monoclonal antibody.

Article Snippet: Bio-Techne (Novus Biologicals) , NBP3-15097 , 230461 , AB_3094817 , recombinant mono , S05-4B6 , rabbit , n/a , Wb, IF.

Techniques: Staining, Immunofluorescence, Concentration Assay, Microscopy, Recombinant