The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000 - 1/10000. Detects a band of approximately 117 kDa (predicted molecular weight: 118 kDa).
1/70. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
1/100 - 1/250.
Activates ubiquitin by first adenylating its C-terminal glycine residue with ATP, and thereafter linking this residue to the side chain of a cysteine residue in E1, yielding an ubiquitin-E1 thioester and free AMP.
Protein modification; protein ubiquitination.
Involvement in disease
Defects in UBA1 are the cause of spinal muscular atrophy X-linked type 2 (SMAX2) [MIM:301830]; also known as X-linked lethal infantile spinal muscular atrophy, distal X-linked arthrogryposis multiplex congenita or X-linked arthrogryposis type 1 (AMCX1). Spinal muscular atrophy refers to a group of neuromuscular disorders characterized by degeneration of the anterior horn cells of the spinal cord, leading to symmetrical muscle weakness and atrophy. SMAX2 is a lethal infantile form presenting with hypotonia, areflexia, and multiple congenital contractures.
Flow cytometric analysis of 2% paraformaldehyde-fixed K562 cells labeling E1 Ubiquitin Activating Enzyme with ab180125 at 1/70 dilution (red) compared to a rabbit IgG control (green), followed by Goat anti rabbit IgG (FITC) secondary antibody at 1/75 dilution.
Immunofluorescent analysis of acetone-fixed Jurkat cells labeling E1 Ubiquitin Activating Enzyme with ab180125 at 1/100 dilution, followed by Goat anti rabbit IgG (Dylight 488) at 1/250 dilution. Counter stained with DAPI (blue).
Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling E1 Ubiquitin Activating Enzyme with ab180125 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.
Western blot - Anti-E1 Ubiquitin Activating Enzyme [EPR14203(B)] antibody (ab180125)
All lanes : Anti-E1 Ubiquitin Activating Enzyme antibody [EPR14203(B)] (ab180125) at 1/5000 dilution
Lane 1 : Fetal brain lysate Lane 2 : K562 cell lysate Lane 3 : HeLa cell lysate Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution