Technical Support Articles
Results (713)
Is it possible to perform CUT&Tag on fixed frozen cells?
Yes. Cell fixation is recommended when the cells are very fragile (easily lysed during experiments), when the cellular signaling pathway is sensitive to the Concanavalin A beads...
Do you observe any differences in CUT&Tag assay performance among different cell lines?
We have performed CUT&Tag on dozens of cultured adherent and suspension cell lines, both human and mouse. So far, we have not noticed any significant difference in performance ...
Can my slides be used for other assays after SignalStar™ Multiplex IHC?
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 ...
Have species been tested if they are not listed as cross-reactive with a CST antibody on the datasheet?
Possibly. CST scientists test all our antibodies on multiple species when feasible, depending on the target protein and the experimental systems available. If a tested species does...
Why are the beads from PTMScan® HS Kits aggregating against the sides of the tube?
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...
Can you help me calculate the bioactivity of my cytokine?
Specific activity (expressed in units/mg) is a function of cytokine or growth factor purity and appropriate protein folding. To determine the specific activity, typically the reado...
Are Smad8 and Smad9 the same protein?
Smad8 and Smad9 are often used interchangeably in the literature to refer to the same protein. However, the official name of the protein is Smad9. Moreover, there ...
Is there a maximum number of cells that can be used in a single CUT&Tag assay?
We have found 100,000 cells per assay to be sufficient for enriching target loci for multiple histone modifications, transcription factors, and transcription cofactors. However, on...
What is the composition of the DNA Elution Buffer #10009?
The composition of our DNA Elution Buffer #10009 is 10 mM Tris-Cl, pH 8.5.
How is the PTMScan® HS Ubiquitin/SUMO Remnant Motif (K-ε-GG) Kit #59322 different from traditional, agarose bead PTMScan®?
The PTMScan® HS Ubiquitin/SUMO Remnant Motif (K-ε-GG) Kit #59322 uses the same antibody as in the traditional PTMScan Ubiquitin Remnant Motif (K-ε-GG) Kit #5562 and PTMScan Pilot U...
Why should I use your recommended protocol or reagents for my IHC experiment?
We have performed a number of comparisons over the years looking at protocol variables such as the retrieval solution, diluent, incubation, and detection methodology. We have seen ...
I don’t see my target of interest in your menu of available antibodies for SignalStar™ Multiplex IHC. Can I still use it in my panel in some way?
SignalStar Multiplex IHC kits and reagents haven’t yet been validated for use with antibodies outside of our menu. We’re in the proces...
Can I autoclave the 6-Tube Magnetic Separation Rack #7017?
No, these racks cannot be autoclaved. Exposing them to high autoclave temperatures may cause the magnets to lose their magnetism. Instead, we suggest submerging or spraying the ...
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. W...
Can I use PTMScan® LysC Protease #84748 for tandem mass tag (TMT) labeling?
We have used LysC in conjunction with trypsin for TMT-labeling-based studies to reduce miscleavages. For TMT-based workflows, we recommend a higher LysC:substrate ratio of 1:100 co...
Which phospho-S6 ribosomal protein (Ser235/236) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) with human samples?
We have several antibodies for detecting the phosphorylation of ribosomal protein S6 (rpS6) at Ser235/236 that are validated for IF-IC with human samples. These are as follows:
What is the expected signal strength for the PTMScan® Control peptides?
We have observed >1E6 peak area for each control peptide when used at the supplied concentration followed by PTMScan and PTMScan HS enrichment...
Do your DYKDDDDK Tag antibodies detect 3X FLAG tag?
Yes, our DYKDDDDK Tag antibodies will detect 3X FLAG tag as long as the FLAG tag sequence is located on the N-terminal or C-terminal end.
Which p44/42 MAPK (Erk1/2) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) with mouse samples?
We have several monoclonal antibodies for studying p44 and p42 mitogen-activated protein kinases (MAPKs), also referred to as Erk2 and Erk1, that are validated in IF-IC with mouse ...
Does CST have a protocol for treating cells with poly(dA:dT)?
The suggested protocol for treating cells with poly(dA:dT) can be found below:
Poly(dA:dT) Treatment Protocol (5 μg/mL final concentra...
Does your Cas9 (7A9-3A3) Mouse mAb #14697 detect the D10A/H840A mutant (dCas9) and the D10A mutant (niCas9)?
Our Cas9 (7A9-3A3) Mouse mAb #14697 was produced with a recombinant protein specific to the amino terminus of Cas9 from Streptococcus pyogenes (UniProt ID #Q99ZW2; https://w...
Do you have a methylene blue dot blot staining protocol?
We have found that the methylene blue staining of the amounts of DNA spotted when performing a DNA dot blot can be tricky and may need optimization.
1. It is very important ...
Is there a method I can use to quantify the amount of protein in my lysate after adding SDS loading buffer?
We use the Pierce 660nm Protein Assay Kit supplemented with the ionic detergent compatibility reagent (IDCR) to determine the protein concentration of lysates prepared in 1X SDS + ...
Does the CUT&Tag assay show a bias toward euchromatin or heterochromatin?
With the appropriate salt concentration in the tagmention buffer, we don't observe bias toward euchromatin or heterochromatin in our assays. In CUT&Tag, the target-specific...
Which S6 ribosomal protein antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with human samples?
We have several antibodies for detecting the phosphorylation of ribosomal protein S6 (rpS6) at Ser235/236 that are validated for IHC-P with human samples. These are S6 Ribosomal Pr...
How do I avoid batch effects and what sample run order should I use for PTMScan® experiments?
To avoid intra-sample batch effects during liquid chromatography with tandem mass spectrometry (LC-MS/MS), we recommend a run order that involves running one technical replicate of...
Which phospho-S6 ribosomal protein (Ser235/236) antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with human samples?
We have several antibodies for detecting the phosphorylation of ribosomal protein S6 (rpS6) at Ser235/236 that are validated for IHC-P with human samples. These are Phospho-S6 Ribo...
How do I determine the concentration of my fractionated samples using the Cell Fractionation Kit #9038?
Since the buffers in the Cell Fractionation Kit #9038 contain SDS, you should use a commercially available ready-to-use, detergent- and reducing agent-compatible assay to quickly m...
Which S6 ribosomal protein antibody do you suggest for immunofluorescence-immunocytochemistry (IF-IC) with mouse samples?
We have several antibodies for detecting ribosomal protein S6 (rpS6) that are validated for IF-IC with mouse samples. These are S6 Ribosomal Protein (5G10) Rabbit mAb #2217 and S6 ...
Does the PathScan® Total Androgen Receptor Sandwich ELISA Kit #12850 recognize the AR-V7 isoform?
Our PathScan® Total Androgen Receptor Sandwich ELISA Kit #12850 recognizes full-length AR protein and is predicted to also recognize the AR-V7 isoform.