affn syk 5a10  (Developmental Studies Hybridoma Bank)

Journal: F1000Research

Article Title: Identification of high-performing antibodies for tyrosine-protein kinase SYK for use in Western Blot, immunoprecipitation and immunofluorescence

doi: 10.12688/f1000research.140456.2

Figure Lengend Snippet: Lysates of THP-1 (WT and SYK KO) were prepared, and 30 μg of protein were processed for Western Blot with the indicated tyrosine-protein kinase SYK 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. An exception was given for antibody GTX633910*, which was titrated to 1/250 as the signal was too weak when following the supplier’s recommendations. Antibody dilutions used: MA1-19332* at 1/1000, 13198** at 1/1000, 80460* at 1/1000, 12358** at 1/1000, GTX633910* at 1/250, MAB7166* at 1/250, NBP1-03250* at 1/500, 66721-1-lg* at 1/2000, AFFN-SYK-5A10* at 1/200, ab3993* at 1/500, ab40781** at 1/1000, ab244701** at 1/1000, ab244968** at 1/1000. Predicted band size: 72 kDa. *Monoclonal antibody, **Recombinant antibody.

Article Snippet: The AFFN-SYK-5A10* antibody was deposited to the Developmental Studies Hybridoma Bank (DSHB) by EU Program Affinomics (DSHB Hybridoma Product AFFN-SYK-5A10).

Techniques: Western Blot, Staining, Membrane, Recombinant

Journal: F1000Research

Article Title: Identification of high-performing antibodies for tyrosine-protein kinase SYK for use in Western Blot, immunoprecipitation and immunofluorescence

doi: 10.12688/f1000research.140456.2

Figure Lengend Snippet: THP-1 WT and SYK 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 indicated tyrosine-protein kinase SYK 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 yellow and magenta dashed line, respectively. Antibody dilutions were chosen according to the recommendations of the antibody supplier. Exceptions were given for antibodies GTX633910* and 66721-1-lg*, which were titrated to 1/2000 and 1/1000, respectively, as the signals were too weak when following the supplier’s recommendations. When the concentration was not indicated by the supplier, which was the case for antibodies ab3993*, ab244701**, and ab244968**, we tested antibodies at 1/1000. At this concentration, the signal from each antibody was in the range of detection of the microscope used. Antibody dilution used: MA1-19332* at 1/1000, 13198** at 1/400, 80460* at 1/400, 12358** at 1/400, GTX633910* at 1/2000, MAB7166* at 1/500, NBP1-03250* at 1/500, 66721-1-lg* at 1/1000, AFFN-SYK-5A10* at 1/60, ab3993* at 1/1000, ab40781** at 1/700, ab244701** at 1/1000, ab244968** at 1/1000. Bars = 10 μm. *Monoclonal antibody, **Recombinant antibody.

Article Snippet: The AFFN-SYK-5A10* antibody was deposited to the Developmental Studies Hybridoma Bank (DSHB) by EU Program Affinomics (DSHB Hybridoma Product AFFN-SYK-5A10).

Techniques: Staining, Concentration Assay, Microscopy, Recombinant

Journal: F1000Research

Article Title: Identification of high-performing antibodies for tyrosine-protein kinase SYK for use in Western Blot, immunoprecipitation and immunofluorescence

doi: 10.12688/f1000research.140456.2

Figure Lengend Snippet: Summary of the Tyrosine-protein kinase SYK antibodies tested.

Article Snippet: The AFFN-SYK-5A10* antibody was deposited to the Developmental Studies Hybridoma Bank (DSHB) by EU Program Affinomics (DSHB Hybridoma Product AFFN-SYK-5A10).

Techniques: Concentration Assay, Recombinant