Technical Support Articles
Results (689)
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-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) with human samples?
We have several monoclonal 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 IF-IC wi...
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...
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?
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 antibodie...
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...
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...
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 ...
Which phospho-Akt (Ser473) antibody do you suggest for Immunofluorescence (Immunocytochemistry) (IF-IC) with human samples?
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:
- Phospho-Akt (Se...
How long after the completion of staining can I wait to image my slides when performing a SignalStar™ Multiplex IHC assay?
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 ...
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 beca...
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 ...
Is PTMScan® compatible with SILAC?
PTMScan® is compatible with the use of stable isotope labeling using amino acids in cell culture, or SILAC. The incorporation of heavy stable iso...
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 ...
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.
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...
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...
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 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...
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...
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...
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...
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 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...
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...
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...
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...
Which p44/42 MAPK (Erk1/2) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) in human samples?
We have several monoclonal antibodies for p44 and p42 mitogen-activated protein kinases (MAPKs), also referred to as Erk2 and Erk1, that are validated for IF-IC with human samples....
Do you have mouse-reactive antibodies available for SignalStar™ Multiplex IHC?
Yes, please select “Mouse” in the first step of the SignalStar Multiplex IH...
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 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...
What are the two bands seen in western blot analysis using ANKRD15 (E9I4I) Rabbit mAb #69953?