The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent dilution. Can be blocked with Human SORBS1 peptide (ab18388). No signal obtained yet but low background observed in human heart and 293 lysates at up to 2 µg/ml.
Use a concentration of 2.5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Plays a role in tyrosine phosphorylation of CBL by linking CBL to the insulin receptor. Required for insulin-stimulated glucose transport. Involved in formation of actin stress fibers and focal adhesions.
Detected in skeletal muscle (at protein level). Widely expressed with highest levels in heart and skeletal muscle.
Contains 3 SH3 domains. Contains 1 SoHo domain.
Cell junction > adherens junction. Cell membrane. Cytoplasm > cytoskeleton. Cell junction > focal adhesion. Colocalizes with actin stress fibers. Also detected at the plasma membrane and in neuronal intranuclear inclusions. Colocalized with PXN at focal adhesions during myogenic differentiation.
Sorbin and SH3 domain-containing protein 1 antibody
Western blot - Anti-SORBS1 antibody (ab4551)
All lanes : Anti-SORBS1 antibody (ab4551)
Lane 1 : Molecular weight marker (Bio-Rad Precision Plus Kaleidoscope standards) Lane 2 : 10 µl Human plasma 1 (600-800 µg protein) Lane 3 : 10 µl Human plasma 2 (600-800 µg protein) Lane 4 : 10 µl Human White Blood cells 1 (30-50 µg protein) Lane 5 : 10 µl Human White Blood cells 2 (30-50 µg protein) Lane 6 : 10 µl HEK293 whole cell lysate (prepared by scrapping 10 cm confluent dish into 4ml of 2x sample buffer (Sigma)
Secondary All lanes : Donkey anti-goat IgG (H+L) conjugated to Alexa-Fluor680 at 1/5000 dilution
ab4551 was tested in in human plasma, human white blood cells and HEK293 cells. Samples were run for 1h at 200 V using the Fisherbrand vertical complete gel unit, and transferred onto nitrocellulose 0.45um (1h, 100V, 4 C) using the Bio-rad Trans blot cell with plate electrodes. Membranes were blocked for 1 h at RT, then incubated overnight in primary antibody (1/200, 1/1000, 1/5000 or minus primary antibody) at 4 degrees C, with shaking. Following washing, membranes were incubated in secondary antibody (1/5000, Alexa-Fluor680 donkey anti-goat IgG, H+L) for 1 h at RT. Antibody binding was detected using LI-COR Bioscience's Odyssey Infrared Imaging system (intensity 5).
The molecular weight of sorbs 1 was calculated using 'Compute pI/MW'from ExPASy tools based on the amino acid sequence according to NP_006425. The band indicated is approximately this predicted molecular weight, and is not present in the minus primary image, scanned at the
ab4551 (2.5 µg/ml) staining SORBS1 in Human Skeletal Muscle by IHC-P. Steamed antigen retrieval in citrate buffer pH6, AP-staining. This image shows spotty staining on muscle fibres in longitudinal section.