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Protein-protein interactions form a network whose structure drives cellular function and whose organization informs all biological inquiry.
Validated antibodies for high content screening from CST Cell Signaling Technology
Through in-house testing, we have found that the magnetic beads from our PTMScan HS kits aggregate and stick to the walls of some specific brands of tubes. Moreover, they cannot be washed off or spun off the sides of the tubes. This is an issue with specific brands of tubes themselves and not an inherent issue with the PTMScan bead product. Therefore, the following brands of polypropylene tubes are NOT recommended with magnetic bead PTMscan HS kits: 1. CellTreat #229441, polypropylene 1.5mL tube, non-sterile, non-pyrogenic2. Denville Scientific (Harvard Bioscience) #C3171, 1.5mL Posi-Lock tubes, polypropyleneThe following brand of microcentrifuge tubes is compatible with magnetic bead PTMscan HS kits:1. USA scientific tubes #1615-5500
Our PTMScan Pan-Methyl Lysine Kit #14809 will enrich for all three forms of methylated lysine: mono, di and tri-methyl lysine. To reduce the complexity of the search space, it is necessary to search for each modification separately, then for all three combined for a total of 4 searches. Therefore, we search for mono-methyl lysine alone as a variable modification, di-methyl lysine alone as a variable modification, tri-methyl lysine alone as a variable modification, then all three forms combined. Each search also has oxidized methionine as a variable modification and the alkylation of cysteine as a static modification. Please note that lysine methylation will often block trypsin cleavage and CST allows for 5 mis-cleavages. Following the 4 independent searches, the results can be combined for a final data set.CST uses the following parameters for the Comet search platform:Pre-cursor mass tolerance (+/-) 50ppmProduct ion match tolerance (+/-) 0.02DaPost-search filter on pre-cursor mass (+…
Webinar | Highly Multiplexed Single Cell Analysis of Tumor Heterogeneity through Time and Space by Mass Cytometry
As a Development Scientist, I focused my attention on research developments in Alzheimer’s Disease (AD), the neurodegenerative disease that affects nearly 2.5 million patients in the U.S. alone.
A scientific resource for the NZF protein domain containing information on structure, function, and domain binding to ubiquitin.
Scientific posters presented by CST scientists at various conferences and trade shows through the years.
Organelle markers for immunofluorescence analysis from CST Cell Signaling Technology
Signal strength by western blot is comparable for our c-Myb (D1B9E) Rabbit mAb #59995 and c-Myb (D2R4Y) Rabbit mAb #12319. However, c-Myb (D1B9E) Rabbit mAb #59995 may show slightly less background.
CST will donate over $10,000 (USD) to the Red Cross to help doctors in China control the spread of the n-coronavirus and more donations are still needed.
Boiling lysates for the detection of mGluR5 should be avoided as it can cause loss of signal. The mGluR5 protein is unstable at higher temperatures and boiling of the lysate will cause it to degrade to the point where it can no longer be detected by our mGluR5 (D6E7B) Rabbit mAb #55920.
It is critical that our Phospho-p53 (Ser15) Antibody #9284 be diluted into 5% w/v non-fat dry milk, 1X TBS, 0.1% Tween® 20 and incubated at 4°C with gentle shaking, overnight. Diluting the antibody in BSA will result in high background on the blot.
A BCA protein assay will likely not provide an accurate concentration for our antibodies formulated in glycerol, due to the presence of BSA and incompatibility with a higher percentage of glycerol.
No matter your research area, we’ve all needed to answer this deceptively easy question: how much of a particular protein is present in my samples?
Chromatin IP (ChIP) Frequently Asked Questions (FAQs) from Cell Signaling Technology, Inc.
A scientific resource for the PAS protein domain containing information on structure, function, and domain binding.
It is difficult to predict when the Fc receptor blocking step is necessary, due to the variable interaction of host species with different antibody isotypes and the interference caused by fluorescent dyes directly conjugated to the antibody Fc domain. The presence of these dyes is often sufficient to disrupt Fc receptor binding. If you are unable to conduct an experiment to assess nonspecific binding, we recommend using the CD16/CD32 (2.4G2) Rat mAb #88280 to prevent any unintended antibody binding to the Fc receptor.
Knockout cell lines and modulating the target's phosphorylation state are experimental controls that can be used to confirm specificity of your cell type.
Lymphoid lineage cells can be characterized using antibodies for cell type-specific markers, allowing cells to be visualized using flow cytometry or IHC.
A scientific resource for the GLUE protein domain containing information on structure, function, and binding to phospholipids in the regulation of vesicle trafficking.
We have no specific tips for using FACS-isolated cells in CUT&RUN experiments. Like a cultured cell suspension, if your isolated cells are in less than 40ul PBS, you can move directly to Concanavalin A bead binding by adding wash buffer for a total of 100ul per reaction. Otherwise, you need to either spin the cells and remove some of the PBS or add wash buffer and Concanavalin A beads proportionally based on the volume of PBS you have in your sorted cell sample. In our experience, the Concanavalin A beads do not bind to cells in the pure PBS environment.
The two dominant bands detected by our TCF4/TCF7L2 antibodies (#2565, #2569, and #2953) represent different splice variants of TCF4/TCF7L2 [see Weise, A. (2010) Nucleic Acids Res 38, 1964-81 (PMID: 20044351; https://pubmed.ncbi.nlm.nih.gov/20044351/)].
ProLong® Gold Antifade Reagent with DAPI #8961 will not freeze solid at -20C. It does, however, become very viscous, and we recommend bringing it to room temperature before pipetting. Freeze-thaw cycles do not appear to affect the efficacy of this product. Additionally, the product is stable for up to 6 months at room temperature, so therefore it is safe to assume it is stable for the same period of time at 4C.
Understanding the microenvironment of disease is critical for targeted therapies. This webcast explores the value of employing image-based methods.
A scientific resource for the DED protein domain containing information on structure, function, and domain binding during apoptosis.
A scientific resource for the CARD protein domain containing information on structure, function, and domain binding during apoptosis.
We have not validated the Anti-rabbit IgG, HRP-linked Antibody #7074 for IHC in-house. Therefore, it is not recommended for this application. We do, however, recommend our SignalStain Boost IHC Detection Reagent (HRP, Rabbit) #8114. It is a highly sensitive, one-step, polymer-based detection reagent that is specific for rabbit IgG. It can be used to visualize targets in both paraffin-embedded and frozen tissue sections and is compatible with all peroxidase-based substrates.
A scientific resource for the FHA protein domain containing information on structure, function, and binding to phospho-serine and phospho-threonine motifs.
Antibody 5G10, which targets S6 ribosomal proteins, can be used as a control for formaldehyde fixation conditions and sample quality in IF experiments.