Log In Sign up
The largest database of trusted experimental protocols

Sc 354x

Manufactured by Santa Cruz Biotechnology
Visit Supplier
Sourced in United States

The Sc-354X is a high-performance laboratory centrifuge designed for a wide range of applications. It features a brushless DC motor and a digital display for accurate speed and time control. The Sc-354X is capable of reaching a maximum speed of 6,000 rpm and can accommodate a variety of rotor options for different sample sizes and types.

Automatically generated - may contain errors

Lab products found in correlation

Chip it express kit, by Active Motif (3 mentions) Bioruptor ucd 200 ultrasound sonicator, by Diagenode (3 mentions) Ab14984, by Abcam (1 mentions) Ab203591, by Abcam (1 mentions) Protein g magnetic beads, by Active Motif (1 mentions) Acetylated histone h3, by Active Motif (1 mentions) Suz12, by Active Motif (1 mentions) Kapa sybr fast qpcr kit, by Roche (1 mentions) Dynabead, by Thermo Fisher Scientific (1 mentions) Protein g dynabead, by Thermo Fisher Scientific (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions)

5 protocols using sc 354x

1

ChIP-qPCR Analysis of ERG Binding

Check if the same lab product or an alternative is used in the 5 most similar protocols
ChIP was performed using the ChIP-IT express kit (Active Motif). Briefly, HUVEC transfected with ERG or control siRNA, and/or treated with Ang1, were crosslinked for 10 min with formaldehyde (to a final concentration of 1%). Chromatin was sheared for five cycles (30 s on, 30 s off) using a Bioruptor UCD-200 ultrasound sonicator (Diagenode), resulting in DNA fragments of 200–1,000 bp in size. Chromatin was immunoprecipitated with 2 μg antibody to ERG (sc-354X, Santa Cruz Biotechnology), or negative control rabbit IgG (PP64, Chemicon, Millipore). Immunoprecipitated DNA was then used as template for quantitative PCR using primers specific for genomic loci. Oligonucleotide sequences are listed in Supplementary Table 1.
Shah A.V., Birdsey G.M., Peghaire C., Pitulescu M.E., Dufton N.P., Yang Y., Weinberg I., Osuna Almagro L., Payne L., Mason J.C., Gerhardt H., Adams R.H, & Randi A.M. (2017). The endothelial transcription factor ERG mediates Angiopoietin-1-dependent control of Notch signalling and vascular stability. Nature Communications, 8, 16002.
+ Open protocol
+ Expand
2

Chromatin Immunoprecipitation Assay for Transcription Factors

Check if the same lab product or an alternative is used in the 5 most similar protocols
Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT Express kit (Active Motif)33 (link), according to the manufacturer’s instructions. In brief, HUVEC were cross-linked for 10 min with 1% formaldehyde. Chromatin was sheared using a Bioruptor UCD-200 ultrasound sonicator (Diagenode) and immunoprecipitated with 3 μg antibody to ERG (sc-354X, Santa Cruz Biotechnology), KLF2 (rabbit, ab203591, Abcam), H3K27Ac (rabbit, 39133, Active Motif) or p300 (mouse, ab14984, Abcam). The respective negative controls were rabbit IgG (PP64, Chemicon, Millipore) and mouse IgG (12- 371, Millipore). Immunoprecipitated DNA was then used as template for quantitative PCR using primers specific for genomic loci and listed in Supplementary Table 5.
Peghaire C., Dufton N.P., Lang M., Salles-Crawley I.I., Ahnström J., Kalna V., Raimondi C., Pericleous C., Inuabasi L., Kiseleva R., Muzykantov V.R., Mason J.C., Birdsey G.M, & Randi A.M. (2019). The transcription factor ERG regulates a low shear stress-induced anti-thrombotic pathway in the microvasculature. Nature Communications, 10, 5014.
+ Open protocol
+ Expand
3

ChIP-qPCR Analysis of ERG and SMAD3 Transcriptional Regulation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Chromatin immunoprecipitation (ChIP) was carried out using ChIP-IT express kit (Active Motif). HUVEC were transfected with ERG or control siRNA (20 nM, for 48 h) or treated with PBS or TGFβ2 (10 ng ml−1, for 30 min), and cross-linked for 10 min with formaldehyde (final concentration of 1%). Chromatin was sheared using a Bioruptor UCD-200 ultrasound sonicator (Diagenode), resulting in DNA fragments of 500–1000 bp in size. Chromatin was immunoprecipitated with 2 μg antibody to ERG (sc-354X, Santa Cruz Biotechnology), SMAD3 (#9523, Cell Signalling) or negative control rabbit IgG (PP64, Chemicon, Millipore) using protein G magnetic beads (Active Motif). Immunoprecipitated DNA was then used as template for qPCR using primers specific for genomic loci. Oligonucleotide sequences are listed in Supplementary Table 5.
Dufton N.P., Peghaire C.R., Osuna-Almagro L., Raimondi C., Kalna V., Chauhan A., Webb G., Yang Y., Birdsey G.M., Lalor P., Mason J.C., Adams D.H, & Randi A.M. (2017). Dynamic regulation of canonical TGFβ signalling by endothelial transcription factor ERG protects from liver fibrogenesis. Nature Communications, 8, 895.
+ Open protocol
+ Expand
4

ChIP-seq Analysis of Protein-DNA Interactions

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell line samples (500,000 per IP) or tissue samples (after homogenization) (25 mg per IP) were exposed to formaldehyde (37%) to cross-link protein–DNA complexes and processed as previously described32 (link),33 (link). Chromatin extracts were immunoprecipitated with antibodies against ERG (sc-354 X, Santa Cruz Biotechnology Inc.), EZH2 (AC22, Active Motif), acetylated histone H3 (Active Motif), tri-methylated histone H3 at lysine 27 (H3K27me3) (Upstate Biotechnology), SUZ12 (Active Motif), AR (#06-680, Millipore), and IgG (Millipore) as control. For ChIP–reChIP experiments, cells were treated and processed as above. Then, before the second round of immunoprecipitation, beads were eluted with 10 mM DTT at 37 °C for 30 min and immunoprecipitation was carried out as described above. ChIP-sequencing was performed using the anti-mERG antibody in VCaP cells.
Quantitative real-time PCR was performed using KAPA SYBR Fast qPCR kit (KAPABiosystems) and primers spanning the region of interest (Supplementary Table 2). The amount of immunoprecipitated DNA was calculated in reference to a standard curve and normalized to input DNA or subsequently to IgG33 (link). For comparison of test and control samples, the amount of DNA was then normalized to the control samples.
Zoma M., Curti L., Shinde D., Albino D., Mitra A., Sgrignani J., Mapelli S.N., Sandrini G., Civenni G., Merulla J., Chiorino G., Kunderfranco P., Cacciatore A., Kokanovic A., Rinaldi A., Cavalli A., Catapano C.V, & Carbone G.M. (2021). EZH2-induced lysine K362 methylation enhances TMPRSS2-ERG oncogenic activity in prostate cancer. Nature Communications, 12, 4147.
+ Open protocol
+ Expand
5

Immunoprecipitation of ERG Transcription Factor

Check if the same lab product or an alternative is used in the 5 most similar protocols
Whole-cell extracts from >3 × 107 MOLT4 cells were prepared using lysis buffer with the following contents: 20 mM HEPES-NaOH (pH 7.8), 300 mM NaCl, 1 mM EDTA, 20% glycerol, 0.5% NP-40, protease inhibitor cocktails (Roche, Basel, Switzerland), 10 mM NaF and 1 mM Na3VO4 (1 ml lysis buffer per 1 × 107 cells). Cleared lysate was incubated overnight with anti-ERG antibody (sc-354x, Santa Cruz, Dallas, TX, USA) at 4 °C (1 μl antibody per 1 × 107 cells) and adsorbed onto protein G Dynabeads (Invitrogen, Carlsbad, CA, USA) for 3 h at 4 °C (20 μl Dynabead suspension per 1 × 107 cells) the next day. The beads were washed with low detergent lysis buffer (20 mM HEPES-NaOH (pH 7.8), 300 mM NaCl, 1 mM EDTA, 20% glycerol and 0.1% NP-40) and the precipitated proteins were subjected to gel electrophoresis or in-solution digestion.
Huang Y., Thoms J., Tursky M., Knezevic K., Beck D., Chandrakanthan V., Suryani S., Olivier J., Boulton A., Glaros E., Thomas S., Lock R., MacKenzie K., Bushweller J., Wong J, & Pimanda J. (2016). MAPK/ERK2 phosphorylates ERG at serine 283 in leukemic cells and promotes stem cell signatures and cell proliferation. Leukemia, 30(7), 1552-1561.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!