Technical Support Articles
Results (713)
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 st...
Do you have mouse-reactive antibodies available for SignalStar™ Multiplex IHC?
Yes, please select “Mouse” in the first step of the SignalStar Multiplex IH...
What is an appropriate positive control to include in this SignalStar™ assay? Are multiple controls necessary?
Any tissue shown to be positive for each marker via chromogenic IHC can serve as a positive control tissue for a SignalStar™ assay. Each target will therefore require a positive co...
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.
What advice do you have for collecting samples of adherent cell lines versus suspension cell lines for CUT&Tag experiments?
For CUT&Tag assays using adherent cell lines, the first step is to detach the cells from the dish. We recommend using trypsin to detach the cells. Alternatively, Accutase Cell ...
How should I store my antibody?
Proper storage of your antibody may vary depending on the product's storage buffer and antibody conjugation. Therefore, please refer to the "Storage" section of the p...
Can CUT&RUN enriched DNA be analyzed by qPCR, or do I have to do NGseq to analyze my data?
Our CUT&RUN Assay Kit #86652 is compatible with downstream q...
How much CUT&RUN DNA is required for NG-seq analysis?
The amount of CUT&RUN DNA required for NG-seq analysis depends on the sensitivity of the DNA library prep kit. The typical yield for CUT&RUN DNA is 0.6 to 6 ng per reaction...
Which total Akt antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with human samples?
We have several monoclonal antibodies for detecting total Akt protein that are validated for IHC-P in human samples. These are as follows:
- Akt (pan) (C67E7) Rabbit mAb #46...
Do you recommend performing size selection of the DNA during the library preparation for CUT&Tag experiments?
No, we don't recommend performing size selection during the library purification for CUT&Tag. Based on our experience and feedback from customers, size selection can signif...
Which method is better for purifying CUT&Tag DNA, DNA affinity columns or a phenol-chloroform DNA extraction using ethanol precipitation?
We recommend using DNA Purification Buffers and Spin Columns (ChIP, CUT&RUN, CUT&Tag) #14209 when purifying your CUT&Tag DNA. Although phenol-chloroform extraction foll...
Why does the material in my new vial of Echinomycin #51434 look different?
Echinomycin #51434 is lyophilized into the vial, and any difference in the appearance of the lyophilized echinomycin is due to a dry coating of the compound in the vial. When you a...
Which phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with human samples?
We have several antibodies for detecting p44 and p42 MAPK when phosphorylated at Thr202 and Tyr204 of Erk1, or Thr185 and Tyr187 of Erk2, that are validated for IHC-P with human sa...
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...
What are the two bands seen in western blot analysis using ANKRD15 (E9I4I) Rabbit mAb #69953?
When using ANKRD15 (E9I4I) Rabbit mAb #69953, two distinct bands are typically observed in western blotting experiments. These bands represent two different isoforms of the ANKRD15...
Which cleaved caspase-3 (Asp175) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) with mouse samples?
We have several antibodies for detecting caspase-3 when cleaved at Asp175 that are validated for IF-IC with mouse samples. These are:
- Cleaved Caspase-3 (Asp175) Antibody #9...
What is the fewest number of cells I can use in a reaction with your CUT&RUN Assay Kit #86652?
We have shown that our CUT&RUN Assay Kit #86652 works with as few as 5,000-10...
Which total Akt antibody do you suggest for immunofluorescence-immunocytochemistry (IF-IC) with mouse samples?
We have several total Akt antibodies that are validated for IF-IC with mouse samples. These are:
- Akt (pan) (C67E7) Rabbit mAb #4691
- Akt (pan) (11E7) Rabbit mAb #468...
Can I use CST antibodies for fluorescent detection in FFPE tissue?
At CST we do not typically validate antibodies using indirect fluorescent detection in formalin fixed, paraffin embedded (FFPE) tissue. If a product is recommended for use in chrom...
What is the expected size distribution of CUT&RUN DNA?
When using antibodies for CUT&RUN against histone modifications, we typically see fragment sizes as small as 150 bp (size of a mono-nucleosome) and quite often see a nucleosoma...
Can the Cell Fractionation Kit #9038 be used for frozen cell pellets?
We have had customers perform this assay using frozen cell pellets, prior to adding any fractionation buffer, and the subsequent fractionation worked fine. Once you start fracti...
What DNA library prep kit do you recommend using for CUT&RUN?
We recommend using our SimpleChIP® ChIP-seq DNA Library Prep Kit for Illum...
What is the most suitable control DNA sample for my CUT&RUN NG-seq experiment?
When performing CUT&RUN with downstream NG-seq analysis, one can use a normal IgG antibody enriched sample as the negative control. However, we have seen and heard from other s...
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...
Why is the PTMScan® LysC Protease #84748 digestion performed at room temperature and not at 37°C?
The digestion is performed at room temperature to minimize carbamylation (also known as carbamoylation) of the sample peptides in the urea buffer, which can interfere with downstre...
What are the specifications for the 96-well plate in the CST ELISA kits?
For the 96-well colorimetric PathScan® and FastScan® ELISA plates, the specification information can be found in the We have several monoclonal antibodies for detecting total Akt protein that are validated for IHC-P with mouse samples. These are as follows:
Which total Akt antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with mouse samples?
Do I need to perform Fc blocking when using rabbit or mouse antibodies in flow cytometry?
Human Fc-gamma Receptor 1a (FCGR1A) exhibits strong cross-reactivity with rabbit IgG, and mouse IgG2a, IgG2c, and IgG3. Of course, mouse Fc-gamma Receptors will react with mouse an...
What is the fewest number of cells I can use in a reaction with the CST CUT&Tag kit?
We have shown that our CUT&Tag Assay Kit #77552 works with as few as 5,000-10,000 cells for histone modifications and 20,000 cells for transcription factors and cofactors.
Can CUT&Tag-enriched DNA be analyzed by qPCR, or do I have to do NGS to analyze my data?
If qPCR analysis is desired, we recommend performing CUT&RUN, as CUT&Tag DNA is not compatible with qPCR. The sample incubation step at 58℃ required for tagmented DNA so...