R960 25

Manufactured by Santa Cruz Biotechnology

The R960-25 is a laboratory centrifuge designed for general-purpose applications. It has a maximum speed of 6,000 rpm and a maximum relative centrifugal force (RCF) of 4,464 x g. The centrifuge accommodates a variety of rotor types and can process sample volumes up to 250 mL.

Automatically generated - may contain errors

Lab products found in correlation

F1804, by Santa Cruz Biotechnology (1 mentions) Cl xposure film, by Thermo Fisher Scientific (1 mentions) Anti h3k9me3, by Abcam (1 mentions) Supersignal west dura extended duration substrate, by Thermo Fisher Scientific (1 mentions) Polyvinylidene difluoride membrane, by Bio-Rad (1 mentions) Mouse anti myc, by Santa Cruz Biotechnology (1 mentions) Rabbit anti histone 3, by Merck Group (1 mentions) Rabbit anti akt, by Cell Signaling Technology (1 mentions) Rabbit anti phospho akt, by Cell Signaling Technology (1 mentions) Mouse anti tubulin, by Merck Group (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Phosphatase inhibitor cocktail, by Roche (1 mentions) Rabbit anti gapdh, by GeneTex (1 mentions) Sc 28364, by Santa Cruz Biotechnology (1 mentions)

3 protocols using r960 25

Immunoblotting Protocol for Protein Analysis

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

Cells were lysed by using lysis buffer (containing protease inhibitor cocktail and PMSF) and the lysate was separated at 12,000g, 20 min. As per the requirement, the concentrations of the protein samples were estimated by Bradford assay. The proteins in the lysate are denatured by boiling for 10 min with 2× Laemmli buffer. The protein samples were loaded onto an SDS–PAGE gel and transferred onto a polyvinylidene difluoride membrane (BioRad). The membrane was blocked with 3% BSA at room temperature overnight, followed by incubation with primary antibodies for 1.5 h at room temperature. The secondary horseradish peroxidase (HRP)-conjugated anti-mouse IgG, HRP-conjugated anti-rabbit IgG or HRP-conjugated anti-goat IgG (1:10,000) antibodies were then added to the membrane at room temperature for 1 h. For signal detection, we used the SuperSignal West Dura Extended Duration Substrate (Thermo Scientific) and captured the blots on CLXposure films (Thermo Scientific). The primary antibodies used were anti-V5 (1:1000, ThermoFisher Scientific #R960-25), anti-ACTIN (1:1000, Santa Cruz #sc-10731), anti-FLAG (1:500, Sigma-Aldrich #F1804), anti-ESET (SETDB1) (1:2000, Santa Cruz #sc-66884), anti-SMC1A (1:2000, Bethyl #A300-055A) and anti-H3K9me3 (1:1000, Abcam #ab8898).

Warrier T., El Farran C., Zeng Y., Ho B.S., Bao Q., Zheng Z.H., Bi X., Ng H.H., Ong D.S., Chu J.J., Sanyal A., Fullwood M.J., Collins J.J., Li H., Xu J, & Loh Y.H. (2022). SETDB1 acts as a topological accessory to Cohesin via an H3K9me3-independent, genomic shunt for regulating cell fates. Nucleic Acids Research, 50(13), 7326-7349.

+ Open protocol

+ Expand

Protein Extraction and Western Blot Analysis

Cited 3 times

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

For protein extraction from S2R+ cells, cell lysates were extracted in lysis buffer (50 mM TrisCl, pH 7.6, 150 mM NaCl, 1 mM EDTA, 1% Triton X-100) with protease inhibitor cocktail (Roche) and phosphatase inhibitor cocktail (Roche). For protein extraction from adult flies, one-day-old flies were collected, and proteins were extracted in lysis buffer with protease inhibitor cocktail (Roche) and phosphatase inhibitor cocktail (Roche). Protein concentrations of the cleared lysate after two times centrifugation (13,000 r.p.m.) at 4 °C were measured, and the same amount of proteins was used for western blot analysis.
For immunostaining of western blots, following antibodies were used: mouse anti-V5 (1:5000, Invitrogen R960-25), mouse anti-Myc (1:1000, Santa Cruz sc-47694), rabbit anti-CCT4 (1:20,000) (this study), rat anti-CCT1 antibody (1:1000, Abcam ab90357), rat anti-Rheb (1:500) [48 (link)], and mouse S6K (1:1000) [40 (link)], rabbit phospho-S6K (1:1000, Cell signaling 9209), rabbit phospho-S6 (1:5000) [49 (link)], rabbit anti-Histone 3 (1:10,000, Millipore 05-928), rabbit anti-GAPDH (1:5000, GeneTex 100118), rabbit anti-Akt (1:1000, Cell Signaling 4691), rabbit anti-phospho-Akt (1:1000, Cell Signaling 4054), mouse anti-Tubulin (1:5000, Sigma T5168), guinea pig anti-TOR (1:2000) [50 (link)], rat anti-TOR (1:2000) [51 (link)].

Kim A.R, & Choi K.W. (2019). TRiC/CCT chaperonins are essential for organ growth by interacting with insulin/TOR signaling in Drosophila. Oncogene, 38(24), 4739-4754.

+ Open protocol

+ Expand

Generation of E2A and HEB knockout ESCs

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

For generation of E2A^−/− ESCs, guide RNAs were designed to target exon 3 of the E2A gene. Targeting was performed on the E2A-V5 cell line to facilitate knockout line validation by loss of V5 signal. For generation of E2A^−/−HEB^−/− double knockout ESCs, exon 9 of the HEB locus was targeted in E2A^−/− knockout cells (clone 9) in order to disrupt both HEBcan and HEBalt splice variants. Guides were designed, assembled as RNPs with recombinant Cas9 and transfected as detailed above. Knockouts were verified by Sanger sequencing of targeted allele and western blot analysis using mouse anti-V5 (1:1000; Thermo Scientific, R960-25) and mouse anti-HEB (1:200; Santa Cruz Biotechnology, sc-28364) antibodies. Experiments presented in the main text were performed using E2A^−/− clone 9 and E2A^−/−HEB^−/− clone 12.

Rao C., Malaguti M., Mason J.O, & Lowell S. (2020). The transcription factor E2A drives neural differentiation in pluripotent cells. Development (Cambridge, England), 147(12), dev184093.

+ 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!