Technical Support Articles
Results (690)
How should I resuspend and aliquot the PTMScan® LysC Protease #84748?
To resuspend the PTMScan® LysC Protease #84748, we recommend allowing the vial to equilibrate at room temperature for a few minutes after removing from the -20C freezer. Next, spin the vial at 1,0...
How does the specificity of the F(ab’)2 fragments differ from the corresponding full-length monoclonal antibodies?
What is the solution formulation of the PTMScan Control peptides?
The solution formulation of the PTMScan Control peptides is 25% acetonitrile, 74.9% water, 0.1% trifluoroacetic acid (V:V).
Why is the concentration of my antibody so low?
At CST, our antibodies are formulated on the basis of their reactivity and performance for each approved application at a recommended dilution. For antibodies with high affinity, optimal performan...
Which cleaved caspase-3 (Asp175) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) with human samples?
We have several antibodies for detecting Cleaved Caspase-3 (Asp175) that are validated for IF-IC with human samples. These are:
- Cleaved Caspase-3 (Asp175) Antibody #9661
- Cleaved Caspas...
Does your CUT&RUN Assay Kit work with isolated nuclei?
Our CUT&RUN Assay Kit is optimized for use with whole cells and works with a large number of cell lines, both adherent and suspension. While it is not typically necessary to pre-isolate cell nucle...
Should digitonin be included in my wash buffers for my entire CUT&Tag experiment?
Yes, digitonin needs to be present in all wash buffers during the entire CUT&Tag experiment. This is because cell membrane permeabilization caused by digitonin is reversible. Removing or lowering ...
Do you have mouse-reactive antibodies available for SignalStar™ Multiplex IHC?
Yes, please select “Mouse” in the first step of the SignalStar Multiplex IHC Panel Builder ...
Should I be concerned that my Concanavalin A beads "clump together" in some of my CUT&RUN experiments?
The Concanavalin A beads can clump together rather easily, especially when the cells become lysed. To avoid this, make sure your cells are healthy and be sure to treat them very gently during the cell...
My items say to store at -20°c but they were shipped in an envelope. Is it ok to use?
Yes, it is safe to use these items. Since 2003, CST has been shipping antibodies at room temperature when possible to reduce waste from packing materials. All CST antibodies are stability tested by ex...
Why can't I add a target to the channel I want when using the SignalStar™ Multiplex IHC Panel Builder?
During our extensive validation process, we found that some targets do not perform optimally with specific fluorophore pairings. When this is the case, the SignalStar Multiplex IHC Panel Builder will ...
When comparing my SignalStar™ staining to the chromogenic staining on a serial section, I see more positive cells. How do I know if this excess staining is correct?
During the course of optimization, we’ve found that fluorescent staining may show higher %-positivity than chromogenic staining. To ensure any excess staining is specific, confirm that the correct sub...
Why do I need to use the three buffers in the Cell Fractionation Kit #9038?
Each buffer in the Cell Fractionation Kit #9038 contains detergents to help isolate different cellular fractions. The Cytoplasmic Isolation Buffer (CIB) contains a non-ionic detergent used as a cell p...
Can I combine antibodies used in a SignalStar™ assay with direct conjugates?
SignalStar Multiplex IHC kits and reagents have been validated for use in combination with direct conjugates. The SignalStar™ Fluorescence Removal Kit Yes, please select “Human” in the first step of the SignalStar Multiplex IHC Panel Builder ... For Imaging Round 1, the staining should show robust signal when imaged up to 8 hours post completion of staining. For Imaging Round 2, imaging should be performed as close to the completion of staini... The specificity of our Raptor (24C12) Rabbit mAb #2280 has been confirmed by siRNA in the following papers: When selecting an antibody for use in CUT&RUN, we recommend starting with a ChIP or ChIP-seq-validated antibody; however, we have found that not all ChIP-validated antibodies work in the CUT&am... Yes, you can use primers from the Illumina Nextera Library preparation system or self-designed primers to amplify the DNA library generated from the CUT&Tag DNA using the CUT&Tag Assay Kit #77... We are confident that the Senescence β-Galactosidase Staining Kit #9860 will continue to perform as expected up to at least 3 freeze/thaw cycles. We recommend aliquoting your reagents into single use ... We have several antibodies for detecting the phosphorylation of Akt at Ser 473 that are validated for IF-IC with human samples. These include the following:
Do you have human-reactive antibodies available for SignalStar™ Multiplex IHC?
How long after the completion of staining can I wait to image my slides when performing a SignalStar™ Multiplex IHC assay?
Can you provide data that confirms the specificity of your Raptor (24C12) Rabbit mAb #2280?
PMID: 28648777 (https://pubmed.ncbi.nlm.nih.gov/28648777/) Figure 2B
PMID: 249...What is the best way to choose an antibody for my CUT&RUN assay if I cannot find a CUT&RUN validated antibody for my target protein?
Can I use primers from the Illumina Nextera Library preparation system or design my own primers to amplify the DNA library generated from the CUT&Tag Assay Kit #77552??
How stable is the Senescence β-Galactosidase Staining Kit #9860 after multiple freeze/thaws?
Which phospho-Akt (Ser473) antibody do you suggest for Immunofluorescence (Immunocytochemistry) (IF-IC) with human samples?
What is the extinction coefficient of my rabbit antibody?
The molar extinction coefficient of rabbit IgG at 280 nm is approximately 210,000 M-1cm-1.
Do I need to optimize the SignalStar™ Multiplex IHC Kits & Reagents for the type of tissue I’m using?
The SignalStar Multiplex IHC kits and reagents have been optimized with respect to fluorophore pairing and order of antibodies. As tissues vary in quality and expression level of biomarkers, increasin...
Which nuclear dyes can be used for In-Cell Western assays in combination with DyLight 680-conjugated secondary antibodies?
When performing In-Cell Western assays with DyLight 680-conjugated secondary antibodies, we recommend using either Hoechst 33342 or DAPI as a nuclear dye. DRAQ5 cannot be used because the emission spe...
Can I reuse PTMScan® antibody beads?
The “classic” PTMScan® kits that use agarose beads and the PTMScan HS kits that use magnetic beads are both designed for a single PTM peptide enrichment. The nature of the acidic elution step, which l...
Which caspase-3 antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with human samples?
We have two antibodies for detecting caspase-3 that are validated for IHC-P with human samples, Caspase-3 Antibody #9662 and Caspase-3 (D3R6Y) Rabbit mAb (IHC Formulated) #14214. We recommend #14214 f...
How are the SignalStar™ Multiplex IHC Kits & Reagents validated?
CST thoroughly validates each antibody available in the PTMScan® is compatible with the use of stable isotope labeling using amino acids in cell culture, or SILAC. The incorporation of heavy stable isotopes has no effect ... CST antibodies undergo thorough validation processes to ensure reliable performance for every approved application. This includes testing different protocol variations to identify the most effective o... SignalStar™ Multiplex IHC Kits & Reagents have not been validated for use in combination with other assays. The SignalStar technology does not destroy the tissue, so it may be possible to perform ...Is PTMScan® compatible with SILAC?
Can I modify the CST-recommended protocol for a given application?
Can my slides be used for other assays after SignalStar™ Multiplex IHC?