Anti ubf1

Manufactured by Santa Cruz Biotechnology

Anti-UBF1 is a laboratory reagent produced by Santa Cruz Biotechnology. It is an antibody that recognizes the UBF1 protein. UBF1 is a transcription factor involved in the regulation of ribosomal RNA genes.

Automatically generated - may contain errors

Lab products found in correlation

Anti s6 ribosomal protein, by Cell Signaling Technology (1 mentions) Nupage system, by Thermo Fisher Scientific (1 mentions) Anti tom20, by Santa Cruz Biotechnology (1 mentions) Anti tfam, by Santa Cruz Biotechnology (1 mentions) Anti fibrillarin, by Santa Cruz Biotechnology (1 mentions) Anti flag m2, by Merck Group (1 mentions) Anti penta his, by Qiagen (1 mentions) Fluorescent mounting medium, by Agilent Technologies (1 mentions) Anti b23, by Merck Group (1 mentions) Anti rnaseh1, by Proteintech (1 mentions)

Close spelling variants of this product

Mouse anti-UBF1 (2 citations)

3 protocols using anti ubf1

Immunoblotting of Subcellular Fractions

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

Immunoblotting was performed with whole cell lysates, fractionated lysates, isolated nuclei or immunoprecipitation samples using the NuPAGE system according to the manufacturer's recommendations (Invitrogen). For preparation of whole cell lysates cells were washed with PBS and lysed in Laemmli lysis buffer (67 mM Tris-HCl pH 6.8, 0.7% SDS). Subcellular fractionation was performed using the ProteoExtrac Subcellular Proteome Extraction Kit (Calbiochem) following the manufacturer's protocol. Immunoprecipitations were performed using standard protocols. The following antibodies were used: anti-penta His (Qiagen), anti-FlagM2 (Sigma), OXPHOS complex I subunit 39 k (Invitrogen), anti-S6 ribosomal protein (Cell Signalling), anti-Tfam, anti-Tom20, anti-Ubf1, anti-Fibrillarin (Santa Cruz Biotechnology). The Alexafluor 680-conjugated secondary antibodies from different species were purchased from Invitrogen. The IRD-800 antibodies were purchased from Rockland. The polyclonal antibody against NOA1 was raised in rabbits by Eurogentec (Liege, Belgium) and detects specifically precursor and mature NOA1 protein (Figure S1 in File S1).

Al-Furoukh N., Kardon J.R., Krüger M., Szibor M., Baker T.A, & Braun T. (2014). NOA1, a Novel ClpXP Substrate, Takes an Unexpected Nuclear Detour Prior to Mitochondrial Import. PLoS ONE, 9(7), e103141.

+ Open protocol

+ Expand

Immunofluorescence Staining of Nuclear Proteins

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

Cells were fixed using 1% formaldehyde for 15 min, washed 3 times with 1X PBS, permeabilized with 0.3% TritonX-100 and then washed again three times with phosphate buffered saline (PBS). Coverslips were blocked using 5% bovine serum albumin for 1 h at RT and transferred to humidified chambers for antibody incubations. Coverslips were incubated with 60 μl of primary antibody for 1 h at RT with concentrations as follows: 1:250 anti-ATXN2 (Sigma HPA021146), 1:250 anti-RNaseH1 (Proteintech, 15606-1-AP), 1:200 anti-B23 (Sigma, B0556) and 1:250 anti-UBF1 (Santa Cruz sc-13125). For all S9.6 R-loop immunofluorescence experiments, 1:500 of the antibody was used. After washing with PBS, cells were incubated with secondary antibody for 1 h in a dark chamber. Following further washing and DAPI staining, coverslips were mounted onto microscope slides using DAKO fluorescent mounting medium and then sealed with nail polish.

Abraham K.J., Chan J.N., Salvi J.S., Ho B., Hall A., Vidya E., Guo R., Killackey S.A., Liu N., Lee J.E., Brown G.W, & Mekhail K. (2016). Intersection of calorie restriction and magnesium in the suppression of genome-destabilizing RNA–DNA hybrids. Nucleic Acids Research, 44(18), 8870-8884.

+ Open protocol

+ Expand

Chromatin Immunoprecipitation of UBF1

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

ChIP was performed as previously described with slight modifications (Neben et al., 2014 (link)). Briefly, cells were cross-linked, and isolated nuclei were sonicated to generate DNA fragments. Chromatin was diluted, and 10% of the supernatant was kept for input. The remaining supernatant was pre-cleared and incubated overnight with 5 μg of antibody (anti-UBF1, Santa Cruz) followed by a 4-hour incubation with Protein G Dynabeads. Protein G bead complexes were washed, and protein-associated chromatin was eluted and cross-linking was reversed. DNA was then purified, precipitated, and quantified by qPCR (rRNA gene UCE Forward 5′-GCGGTTTTCTTTCATTGACC-3′ and Reverse 5′-GTATGACTTCCAGGCGTCGT-3′; 5′-ETS Forward 5′-GCTTGTTTCTCCCGATTGC-3′ and Reverse 5′-ACTTTTCTCAGTGGTTCGCG-3′; and 28S Forward 5′-CGCCGGTGAAATACCACTAC-3′ and Reverse 5′-GTGTCCTAAGGCGAGCTCAG-3′). ChIP enrichment was determined as percentage of input.

Neben C.L., Lay F.D., Mao X., Tuzon C.T, & Merrill A.E. (2016). Ribosome biogenesis is dynamically regulated during osteoblast differentiation. Gene, 612, 29-35.

+ 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!