PRODUCT CODE: ET1607-5

Recombinant Phospho p95/NBS1 (S343) Monoclonal Antibody (ET1607-5)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

REACTIVITY

  • Human

ICC staining of Phospho-p95/NBS1 (S343) in PC-3M cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1607-5, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-p95/NBS1 (S343) in PC-3M cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1607-5, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-Phospho-p95/NBS1 (S343) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1607-5, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC staining of Phospho-p95/NBS1 (S343) in PC-3M cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1607-5, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant Phospho p95/NBS1 (S343) Monoclonal Antibody (ET1607-5)

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding ser343 of human p95/nbs1.

Host

Rabbit

Modification

Phospho

Modification Site

S343

Positive Control

PC-3M, mouse testis tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SY0215

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

95 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

Phospho-p95/NBS1 (S343)

SYNONYMS

AT V1 antibody; AT V2 antibody; ATV antibody; Cell cycle regulatory protein p95 antibody; FLJ10155 antibody; MGC87362 antibody; Nbn antibody; NBN_HUMAN antibody; NBS 1 antibody; NBS antibody; NBS1 antibody; Nibrin antibody; Nijmegen breakage syndrome 1 (nibrin) antibody; Nijmegen breakage syndrome antibody; Nijmegen breakage syndrome protein 1 antibody; p95 antibody; p95 protein of the MRE11/RAD50 complex antibody

TISSUE SPECIFICITY

Ubiquitous. Expressed at high levels in testis.

POST-TRANSLATIONAL MODIFICATION

Phosphorylated by ATM in response of ionizing radiation, and such phosphorylation is responsible intra-S phase checkpoint control and telomere maintenance.

SUBCELLULAR LOCATION

Nucleus, PML body, telomere, Chromosome.

FUNCTION

Nijmegen breakage syndrome (NBS) is characterized by extreme radiation sensitivity, chromosomal instability and cancer. These phenotypes are similar to those of ataxia telangiectasia mutated (ATM) disease, where there is a deficiency in a protein kinase that is activated by DNA damage, indicating that the NBS1 (Nibrin) and ATM proteins may participate in common pathways. Nibrin is specifically phosphorylated in response to gamma-radiation, ultraviolet light and exposure to hydroxyurea. The phosphorylation of Nibrin requires catalytically active ATM. ATM physically interacts with and phosphorylates Nibrin on Serine 343 both in vitro and in vivo. Serine 343 is phosphorylated in vitro by ATM and the modification of this residue in vivo is essential for the cellular response to DNA damage. This response includes S-phase checkpoint activation, formation of the NBS1/Mrel1/Rad50 nuclear foci and rescue of hypersensitivity to ionizing radiation.