Sc 13125

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Sc-13125 is a lab equipment product offered by Santa Cruz Biotechnology. It is designed for general laboratory use. The core function of Sc-13125 is to [description not available].

Automatically generated - may contain errors

Lab products found in correlation

8 protocols using sc 13125

Comprehensive Immunoblotting Antibody Panel

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following antibodies were used: rabbit polyclonal antibody against phospho S139 gamma H2A.X (ab2893, Abcam), rabbit monoclonal antibody against alpha-tubulin (ab52866, Abcam), mouse monoclonal antibody against UBF (sc13125, Santa Cruz), RPA194 (sc48385, Santa Cruz), phospho S139 gamma H2A.X antibody (ab26350, Abcam), Anti-BrdU antibody (B8434, Sigma), sheep polyclonal antibody against BrdU (ab1893, Abcam), Alexa 488-conjugated goat anti-mouse IgG (A11029, Invitrogen), Alexa 568-conjugated goat anti-rabbit IgG (A11011, Invitrogen), DyLight 488 goat anti-mouse IgG (ab96879, Abcam), isotype mouse IgG antibody (ab91353, Abcam), alkaline phosphate-conjugated rabbit IgG (ZB5305, ZSGB) and HRP goat anti-mouse IgG (A0208, Beyotime).

Xie Q., Li C., Song X., Wu L., Jiang Q., Qiu Z., Cao H., Yu K., Wan C., Li J., Yang F., Huang Z., niu B., Jiang Z, & Zhang T. (2016). Folate deficiency facilitates recruitment of upstream binding factor to hot spots of DNA double-strand breaks of rRNA genes and promotes its transcription. Nucleic Acids Research, 45(5), 2472-2489.

+ Open protocol

+ Expand

Immunofluorescence and Western Blotting Protocols

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies used in this study are as follows: UBF (F-9) mouse monoclonal antibody used for immuno-FISH (sc-13125; Santa Cruz Biotechnology), UBF (M01) mouse monoclonal antibody used for immunofluorescence and Western blotting (clone 6B6; Abnova), c-Myc rabbit polyclonal antibody (5605; Cell Signaling Technology), nucleolin rabbit polyclonal antibody (ab70493; Abcam), GAPDH (D16H11) rabbit mAb (5174; Cell Signaling Technology), RAD21 (D213) antibody (4321; Cell Signaling Technology), β-Actin (8H10D10) mouse mAb (3700; Cell Signaling Technology), Ki-67 (8D5) mouse mAb (9449; Cell Signaling Technology), and α-tubulin antibody (ab15246; 1:500; Abcam). Secondary antibodies for immunofluorescence (Alexa Fluor 488 and 594 conjugates) were obtained from Life Technologies and used at 1:500 dilution. Secondary HRP-conjugated antibodies for Western blotting were from Cell Signaling Technology and typically used at 1:5,000 dilution.

Potapova T.A., Unruh J.R., Yu Z., Rancati G., Li H., Stampfer M.R, & Gerton J.L. (2019). Superresolution microscopy reveals linkages between ribosomal DNA on heterologous chromosomes. The Journal of Cell Biology, 218(8), 2492-2513.

+ Open protocol

+ Expand

GFP, SEC23B, and UBF Immunoprecipitation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were pelleted and lysed with M-PER (Thermo Scientific Pierce) supplemented with a cocktail of protease and phosphatase inhibitors (Sigma-Aldrich). Protein lysates were collected by centrifugation at 13 000 rpm for 10 min at 4°C and pre-cleared by incubation with Thermo Protein A/G Dynabeads for 3 h at 4°C on a rotator. Pre-cleared protein lysates were quantified with the BCA Protein Assay Kit (Thermo Scientific Pierce), and 1 mg/ml lysates were prepared. We used anti-GFP (Abcam ab290), anti-SEC23B (Abcam ab151258) and anti-UBF (Santa Cruz sc-13125) antibodies for pull-down and immunoblotting at the recommended dilutions. Cell lysates were separated by SDS-PAGE and transferred onto nitrocellulose membranes. Blots were scanned digitally and quantified using the Odyssey Infrared Imaging System (Li-Cor Biosciences).

Yehia L., Jindal S., Komar A.A, & Eng C. (2018). Non-canonical role of cancer-associated mutant SEC23B in the ribosome biogenesis pathway. Human Molecular Genetics, 27(18), 3154-3164.

+ Open protocol

+ Expand

Antibody Detection and Localization in Western Blotting and Immunofluorescence

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary antibodies used for western blotting were as follows: mouse anti-β-tubulin (T4026, Sigma, 1:6,000), mouse anti-PICH (Ab57434, Abcam, 1:500), goat anti-TopoIIα (sc5347, Santa Cruz, 1:1,000), mouse anti-Actin (A3853, Sigma, 1:6,000), mouse anti-GFP (11814460001, Roche, 1:1,000). Primary antibodies used for indirect immunofluorescence microscopy were as follows: mouse anti-PICH (04-1540, Millipore, 1:100), rabbit anti-PICH (8886S, Cell Signaling, 1:100), guinea pig anti-PICH (in-house, 1:400) mouse anti-β-tubulin (T4026, Sigma, 1:600), rabbit anti-CENPA (a kind gift from Professor Tatsuo Fukagawa published in ref. 30 (link), 1:100), mouse anti-UBF (sc-13125, Santa Cruz, 1:250).

Nielsen C.F., Huttner D., Bizard A.H., Hirano S., Li T.N., Palmai-Pallag T., Bjerregaard V.A., Liu Y., Nigg E.A., Wang L.H, & Hickson I.D. (2015). PICH promotes sister chromatid disjunction and co-operates with topoisomerase II in mitosis. Nature Communications, 6, 8962.

+ Open protocol

+ Expand

Immunofluorescent Localization of UBF

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were seeded on coverslips, washed with PBS, and fixed by incubation in 4% formaldehyde for 20 min at room temperature. The cells were then washed twice in PBS and permeabilized in PBS containing 0.2% Triton x100 and 1% BSA at room temperature for 30 min. Cells were probed with primary antibody to UBF (sc-13125, Santa Cruz), and conjugated secondary anti-mouse Alexa Flour 647 (cat No. 705-605-147, Jackson Labs) antibodies were then applied for detection. To stain the nuclei, the cells were incubated for 30 min with 0.05 μg/ml Hoechst dye (Sigma). Cells were examined and photographed under a confocal laser-scanning microscope (Zeiss LSM 510 META, or Leica STED Live Imaging).

Gelgor A., Gam ze Letova C., Yegorov Y., Kalt I, & Sarid R. (2018). Nucleolar stress enhances lytic reactivation of the Kaposi’s sarcoma-associated herpesvirus. Oncotarget, 9(17), 13822-13833.

+ Open protocol

+ Expand

Histone Modification Profiling by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols

For Western blot analysis of histone modifications and associated proteins, the following antibodies were used: monoclonal anti-HP1α (Millipore 05-689, 1:1000), monoclonal anti-HP1β (Serotec MCA1946, 1:1000, CST 8676S, 1:1000), monoclonal anti-HP1γ (Euromedex clone 2MOD-1G6, 1:000), monoclonal anti-b-tubulin (Sigma-Aldrich, 1:000), polyclonal anti-H3K27me3 (Active Motif 39156, 1:2000), polyclonal anti-H3K9me3 (Invitrogen 49-1008, 1:1000), polyclonal anti-H3 (Abcam ab1791, 1:2000), polyclonal anti-H4K20me3 (IMP 0083, 1:2000), polyclonal anti-H4 (Abcam ab10158, 1:4000), polyclonal anti-Myc (Abcam ab9132, 1:1000), monoclonal anti-GFP (Roche 11814460001, 1:1000). Antibodies used for IF stainings and transmission immuno-electron microscopy were: Monoclonal anti-NPM1 (Invitrogen 32–5200, 1:200), monoclonal anti-FBL (CST 2639 1:200), monoclonal anti-UBF1 (Santa Cruz sc-13125, 1:200), polyclonal anti-H3K9me3 (Active motif 39161, 1:1000), monoclonal anti-HP1β (Serotec MCA1946, 1:500, CST 8676S, 1:500), monoclonal anti-HP1α (Millipore 05–689, 1:500), monoclonal anti-HP1γ (Euromedex 2MOD-1G6-AS, 1:1000), polyclonal anti-H4K20me3 (IMP 0083, 1:2000), polyclonal anti-H3K27me3 (Active motif 39156, 1:500), monoclonal anti-H2AK119ub1 (Upstate 05-678, 1:50), polyclonal anti-Myc (Abcam ab9132, 1:500).

Ballmer D., Tardat M., Ortiz R., Graff-Meyer A., Ozonov E.A., Genoud C., Peters A.H, & Fanourgakis G. (2022). HP1 proteins regulate nucleolar structure and function by secluding pericentromeric constitutive heterochromatin. Nucleic Acids Research, 51(1), 117-143.

+ Open protocol

+ Expand

Immunofluorescence Analysis of Nucleolar Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

The cells were cultured in DMEM in 10% FBS or 0.2% FBS for 18h on Nuc Lab-Tek II chamber slide (Sigma-Aldrich). Cells were washed with PBS, fixed for 15 min with 4% paraformaldehyde (Sigma-Aldrich St. Louis, Mo, USA), washed with PBS, permeabilized with 0.5% Triton X-100 for 10 min and blocked for 40 min with 0.2% gelatin. All incubations with primary antibodies were performed in PBS-Triton X-100 overnight at 4°C. The following primary antibodies were used for immunofluorescence: rabbit polyclonal against N-terminal of EGR1 (4153, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal antibody anti-fibrillarin (ab4566, Abcam, Cambridge, MA, USA), mouse monoclonal antibody anti-B23 (ab10530, Abcam) and mouse monoclonal anti-UBF antibody (sc-13125, Santa Cruz Biotechnology, Dallas, TX, USA). Primary antibodies were diluted at 1∶200. Cells were than washed 3 × 5 min and incubated with secondary antibodies, Alexa Fluor mouse 594 and Alexa Fluor rabbit 488 diluted 1∶1000. Confocal analysis was performed with a Leica SP2. Transcription of rDNA genes was inhibited by supplementing the medium with 0.04 µg/ml actinomycin D (Sigma-Aldrich St. Louis, Mo, USA) for 1h at 37°C the cells were than fixed and stained. The analysis was performed by immunofluorescence microscopy (LEICA DM4000B).

Ponti D., Bellenchi G.C., Puca R., Bastianelli D., Maroder M., Ragona G., Roussel P., Thiry M., Mercola D, & Calogero A. (2014). The Transcription Factor EGR1 Localizes to the Nucleolus and Is Linked to Suppression of Ribosomal Precursor Synthesis. PLoS ONE, 9(5), e96037.

+ Open protocol

+ Expand

Immunohistochemical Analysis of UBTF-TDs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Photomicrographs of bone marrow core biopsy of 4 cases with UBTF-TDs and 2 cases with either KMT2A rearrangement or NPM1c mutation using anti-UBF (sc-13125, Santa Cruz Biotechnology, clone: F-9, RRID: AB_671403, 1:500 dilution). All images are at equal magnification (60×).

Umeda M., Ma J., Huang B.J., Hagiwara K., Westover T., Abdelhamed S., Barajas J.M., Thomas ME I.I.I., Walsh M.P., Song G., Tian L., Liu Y., Chen X., Kolekar P., Tran Q., Foy S.G., Maciaszek J.L., Kleist A.B., Leonti A.R., Ju B., Easton J., Wu H., Valentine V., Valentine M.B., Liu Y.C., Ries R.E., Smith J.L., Parganas E., Iacobucci I., Hiltenbrand R., Miller J., Myers J.R., Rampersaud E., Rahbarinia D., Rusch M., Wu G., Inaba H., Wang Y.C., Alonzo T.A., Downing J.R., Mullighan C.G., Pounds S., Babu M.M., Zhang J., Rubnitz J.E., Meshinchi S., Ma X, & Klco J.M. (2022). Integrated Genomic Analysis Identifies UBTF Tandem Duplications as a Recurrent Lesion in Pediatric Acute Myeloid Leukemia. Blood Cancer Discovery, 3(3), 194-207.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!