The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/100 - 1/500. Antigen retrieval is not essential but may optimise staining. The use of citrate buffer will enhance staining. Likely to work in frozen sections.
Use at an assay dependent concentration.
Component of the MRN complex, which plays a central role in double-strand break (DSB) repair, DNA recombination, maintenance of telomere integrity and meiosis. The complex possesses single-strand endonuclease activity and double-strand-specific 3'-5' exonuclease activity, which are provided by MRE11A. RAD50 may be required to bind DNA ends and hold them in close proximity. This could facilitate searches for short or long regions of sequence homology in the recombining DNA templates, and may also stimulate the activity of DNA ligases and/or restrict the nuclease activity of MRE11A to prevent nucleolytic degradation past a given point. The complex may also be required for DNA damage signaling via activation of the ATM kinase. In telomeres the MRN complex may modulate t-loop formation.
Expressed at very low level in most tissues, except in testis where it is expressed at higher level. Expressed in fibroblasts.
Involvement in disease
Defects in RAD50 are the cause of Nijmegen breakage syndrome-like disorder (NBSLD) [MIM:613078]; also called NBS-like disorder or RAD50 deficiency. NBSLD is a disorder similar to Nijmegen breakage syndrome and characterized by chromosomal instability, radiation sensitivity, microcephaly, growth retardation, short stature and bird-like face. Immunodeficiency is absent.
Belongs to the SMC family. RAD50 subfamily. Contains 1 zinc-hook domain.
The zinc-hook, which separates the large intramolecular coiled coil regions, contains 2 Cys residues that coordinate one molecule of zinc with the help of the 2 Cys residues of the zinc-hook of another RAD50 molecule, thereby forming a V-shaped homodimer. The two heads of the homodimer, which constitute the ATP-binding domain, interact with the MRE11A homodimer.
Phosphorylated upon DNA damage, probably by ATM or ATR.
Nucleus. Chromosome > telomere. Localizes to discrete nuclear foci after treatment with genotoxic agents.
Detection of Human Rad50 by Immunohistochemistry. Sample: FFPE section of Human larynx squamous cell carcinoma.Antibody: ab84397 used at a dilution of 1/250. Detection: DAB staining using anti-Rabbit IHC antibody at a dilution of 1/100.
Detection of Mouse Rad50 by Immunohistochemistry. Sample: FFPE section of Mouse squamous cell carcinoma.Antibody: ab84397 used at a dilution of 1/250. Detection: DAB staining using anti-Rabbit IHC antibody at a dilution of 1/100.
Immunohistochemistry (Frozen sections) - Anti-Rad50 antibody (ab84397)This image is courtesy of an Abreview submitted by Anna Lekawa
ab84397 staining Rad50 in Mouse retinal tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at room temperature. Samples were incubated with the undiluted primary antibody in blocking solution for 16 hours at 4°C. An undiluted Biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.