MA5-32284

antibody from Invitrogen Antibodies

Targeting: RAD18 RNF73

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-32284

Provider

Invitrogen Antibodies

Product name

RAD18 Recombinant Rabbit Monoclonal Antibody (SN66-01)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Antibody clone number

SN66-01

Vial size

100 µL

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

A positive feedback loop: RAD18-YAP-TGF-β between triple-negative breast cancer and macrophages regulates cancer stemness and progression.

Yan X, He Y, Yang S, Zeng T, Hua Y, Bao S, Yang F, Duan N, Sun C, Liang Y, Fu Z, Huang X, Li W, Yin Y
Cell death discovery 2022 Apr 12;8(1):196

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemical analysis of RAD18 in Hela cells using a RAD18 Monoclonal antibody (Product # MA5-32284) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemical analysis of RAD18 in NIH/3T3 cells using a RAD18 Monoclonal antibody (Product # MA5-32284) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemical analysis of RAD18 in 293 cells using a RAD18 Monoclonal antibody (Product # MA5-32284) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow Cytometric analysis of RAD18 in Hela cells using a RAD18 Monoclonal Antibody (Product # MA5-32284) at a dilution of 1:50, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 2 RAD18 promotes TNBC cell proliferation and maintains the stemness of TNBC in vitro. RAD18 increases TNBC cell proliferation and maintains the CSC stemness in vitro. A RAD18 protein levels in TNBC cells transfected with RAD18 shRNA lentivirus were determined by western blotting. B Cells were seeded in a 96-well plate and CCK-8 assay was used to detect cell proliferation at indicated hours (24, 48, 72, 96, and 120 h). C colony formation assay was performed to examine cell proliferation. Cells were seeded in six-well plates at a density of 2000 cells per well and cultivated for 2w. D An EdU assay was performed to determine the proliferation of different groups of TNBC cells. E Cell apoptosis in shNC and shRAD18 TNBC cells were assessed by Annexin V/7-AAD flow cytometric analysis. F Representative flow cytometric analysis of CD44+/CD24- BCSC population percentages in shNC and shRAD18 TNBC cells. G The tumor sphere formation in TNBC cells treated as indicated (200x). The quantification of mammospheres were counted using a microscope with size >=50 um. H , I mRNA and protein levels of stemness-related signature genes (CD44, OCT4, SOX2, Nanog). GAPDH was analyzed as a loading control. * p < 0.05, ** p < 0.01, *** p < 0.001. All experiments were performed independently at least three times.