mAbs Against MOUSE Protein
p19arf
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | Rat monoclonal |
| CLONE NAME: | PIL346C |
| PROTEIN: | Cdkn2a |
| PROTEIN WEB: | https://www.uniprot.org/uniprot/Q64364 |
| ANTIGEN USED: | His-MBP-mp19arf |
| FUSION PARTNER: | NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | IgG2b |
| SPECIES REACTIVITY: | mouse |
| PREPARATION AND STORAGE: | Aliquot and store at 4C. Do not freeze |
Description
Capable of inducing cell cycle arrest in G1 and G2 phases. Acts as a tumor suppressor. Binds to MDM2 and blocks its nucleocytoplasmic shuttling by sequestering it in the nucleolus. This inhibits the oncogenic action of MDM2 by blocking MDM2-induced degradation of p53 and enhancing p53-dependent transactivation and apoptosis. Also induces G2 arrest and apoptosis in a p53-independent manner by preventing the activation of cyclin B1/CDC2 complexes. Binds to BCL6 and down-regulates BCL6-induced transcriptional repression. Binds to E2F1 and MYC and blocks their transcriptional activator activity but has no effect on MYC transcriptional repression. Binds to TOP1/TOPOI and stimulates its activity. This complex binds to rRNA gene promoters and may play a role in rRNA transcription and/or maturation. Interacts with NPM1/B23 and promotes its polyubiquitination and degradation, thus inhibiting rRNA processing. Interacts with COMMD1 and promotes its 'Lys63'-linked polyubiquitination (By similarity). Interacts with UBE2I/UBC9 and enhances sumoylation of a number of its binding partners including MDM2 and E2F1. Binds to HUWE1 and represses its ubiquitin ligase activity. May play a role in controlling cell proliferation and apoptosis during mammary gland development. Isoform smARF may be involved in regulation of autophagy and caspase-independent cell death; the short-lived mitochondrial isoform is stabilized by C1QBP.
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | PIL346C | Neat | supernatant | / | |||||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | PIL346C | 1:10 | supernatant | Fibrosarcoma / Liver | nuclear | ||||
| Immunofluorescence | |||||||||
Enlarge image
- PIL346C mAb is able to detect mouse p19 protein in immunocytochemistry
- To confirm that PIL346C mAb recognizes human p19 protein, immunocytochemistry on frozen cytospin preparations of p19 and p16 expressed in HEK293 cells was performed. Anti-V5 confirmed the transfection efficiency.
Enlarge image
- PIL346C mAb can be used to detect mouse p19 protein by immunohistochemistry
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | PIL346C | Neat | supernatant | / | 19kDa | 19kDa | Available | |
| Immunoprecipitation | ||||||||
Enlarge image
- PIL346C mAb is able to detect mouse p19 protein by WB
- LANES
Lane 1 HEK-B-MYB (20ug) (-)
Lane 2 HEK-V5-mP19 (20ug) (+)
Lane 3 Mefs RASV12 (50ug) (+)
Lane 4 Mef (50ug) (-)
Beta actin was used as loading control.