Ab104306

Manufactured by Abcam

Ab104306 is a primary antibody product from Abcam. It is a monoclonal antibody that recognizes a specific target protein. The core function of this product is to serve as a research tool for the detection and study of the target protein in various experimental applications.

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Lab products found in correlation

Alexa fluor 647 goat anti mouse, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (1 mentions) Ab133534, by Abcam (1 mentions) Ab26350, by Abcam (1 mentions) Dm1000 led, by Leica (1 mentions) Nucblue live readyprobes reagent, by Thermo Fisher Scientific (1 mentions) Lsm 780 inverse laser scanning microscope, by Zeiss (1 mentions) A32731, by Thermo Fisher Scientific (1 mentions) Lsm t pmt detector, by Zeiss (1 mentions) A11032, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)

2 protocols using ab104306

Quantifying RAD51C Variant DNA Damage Response

Cited 1 time

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Stable landing pad U2OS cells expressing RAD51C variants were irradiated with 5 Gy (CellRad, Precision X-Ray) and fixed with 4% paraformaldehyde 4 hours postirradiation. Cells were incubated for 1 hour with anti-RAD51 (polyclonal; Bioacademia #70–001) and anti-geminin (monoclonal; Abcam #ab104306) antibodies (32 (link)), and secondary antibodies [Alexa Fluor 488 goat anti-rabbit (Thermo Fisher Scientific, #A-11008) and Alexa Fluor 647 goat anti-mouse (Thermo Fisher Scientific #A-21235)] and stained with DAPI. Z-stack microscopy images from a 50x water immersion objective were automatically acquired for 1,000 geminin-positive (S/G₂ phase) cells per variant in a Cell discoverer 7 instrument (Zeiss). Images were processed and analyzed using ZEN (blue edition) Celldiscoverer 3.2 (Zeiss) software. Each RAD51C variant was assessed in triplicate, unless denoted otherwise.

Hu C., Nagaraj A.B., Shimelis H., Montalban G., Lee K.Y., Huang H., Lumby C.A., Na J., Susswein L.R., Roberts M.E., Marshall M.L., Hiraki S., LaDuca H., Chao E., Yussuf A., Pesaran T., Neuhausen S.L., Haiman C.A., Kraft P., Lindstrom S., Palmer J.R., Teras L.R., Vachon C.M., Yao S., Ong I., Nathanson K.L., Weitzel J.N., Boddicker N., Gnanaolivu R., Polley E.C., Mer G., Cui G., Karam R., Richardson M.E., Domchek S.M., Yadav S., Hruska K.S., Dolinsky J., Weroha S.J., Hart S.N., Simard J., Masson J.Y., Pang Y.P, & Couch F.J. (2023). Functional and Clinical Characterization of Variants of Uncertain Significance Identifies a Hotspot for Inactivating Missense Variants in RAD51C. Cancer Research, 83(15), 2557-2571.

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Evaluating DNA Damage and Repair Dynamics

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Cells were treated with DMSO or 10 μmol/L rucaparib for 24, 48, or 72 hours. Cells were fixed with either 4% paraformaldehyde or methanol, permeabilized with 0.2% TritonX-100, blocked with blocking buffer (1% BSA, 2% FCS or 1% BSA, 3% milk, 2% goat serum in PBS) and incubated with rabbit anti-RAD51 (ab133534 1:250; ab11055 1:400; Abcam) and either mouse anti-Geminin (ab104306 1:100; Abcam) or mouse anti-γH2AX (ab26350 1:400; Abcam) antibodies. For RAD51 foci formation with geminin staining, anti-rabbit 647, and anti-mouse 546 Alexa Fluor secondary antibodies (1:800; Invitrogen Molecular Probes) were used. Nuclei were counterstained with Hoechst 33342 (NucBlue Live ReadyProbes Reagent; Ther-mofisher Scientific). Cells were imaged using an LSM 780 inverse laser scanning microscope (Zeiss) and captured with a LSM T-PMT detector (Zeiss). At least 194 cells from four fields of view and two independent experiments were counted. Cells with ≥5 RAD51 foci/nucleus were scored using CellProfiler (version 2.2.0, Broad Institute). For RAD51 and γH2AX foci formation assay, anti-rabbit 488 and anti-mouse 594 Alexa Fluor secondary antibodies (A32731 1:500; A-11032 1:500; Invitrogen Molecular Probes) were used. Nuclei were counterstained with DAPI (Sigma). Cells were imaged using Leica DM 1000 LED at 40×.

Kondrashova O., Nguyen M., Shield-Artin K., Tinker A.V., Teng N.N., Harrell M.I., Kuiper M.J., Ho G.Y., Barker H., Jasin M., Prakash R., Kass E.M., Sullivan M.R., Brunette G.J., Bernstein K.A., Coleman R.L., Floquet A., Friedlander M., Kichenadasse G., O'Malley D.M., Oza A., Sun J., Robillard L., Maloney L., Giordano H., Wakefield M.J., Kaufmann S.H., Simmons A.D., Harding T.C., Raponi M., McNeish I.A., Swisher E.M., Lin K.K, & Scott C.L. (2017). Secondary Somatic Mutations Restoring RAD51C and RAD51D Associated with Acquired Resistance to the PARP Inhibitor Rucaparib in High-Grade Ovarian Carcinoma. Cancer discovery, 7(9), 984-998.

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