Technical Support Articles
Results (694)
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...
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 ...
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 completion of staini...
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 antibodies work in the CUT&am...
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.
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 (Ser473) (D9E) XP® Rabb...
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 SignalStar Multiplex IHC kits and reagents haven’t yet been validated for use with antibodies outside of our menu. We’re in the process of developing cust... 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... 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... 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 control, which may som... 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... 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 ... 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 ... 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 rack with 10% bleach... 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 protein:
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?
Which nuclear dyes can be used for In-Cell Western assays in combination with DyLight 680-conjugated secondary antibodies?
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?
What is an appropriate positive control to include in this SignalStar™ assay? Are multiple controls necessary?
Which caspase-3 antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with human samples?
Can my slides be used for other assays after SignalStar™ Multiplex IHC?
Is PTMScan® compatible with SILAC?
Can I autoclave the 6-Tube Magnetic Separation Rack #7017?
What are the two bands seen in western blot analysis using ANKRD15 (E9I4I) Rabbit mAb #69953?
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 nucleosomal ladder of DNA frag...
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 #4685
- Akt Antibo...
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 not show cross-reac...
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 product webpage or to...
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 #9661
- Cleaved ...
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 chromogenic IHC, denoted ...
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 5,000-10,000 cells for histo...
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 qPCR analysis. The pr...
What DNA library prep kit do you recommend using for CUT&RUN?
We recommend using our SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #567...
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 downstream PTMScan enrichmen...
How did you determine that a NG-seq sequencing depth of 3 to 5 million sequencing reads was sufficient for CUT&RUN analysis?
This decision was based on our analysis of hundreds of CUT&RUN samples. We found that a sequencing depth lower than 3 million generated too few peaks and that the number of peaks increased with de...
Which total Akt antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with mouse samples?
We have several monoclonal antibodies for detecting total Akt protein that are validated for IHC-P with mouse samples. These are as follows:
- Akt (pan) (C67E7) Rabbit mAb #4691
- Akt (pa...
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. Our 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 scientists that norma...What is the most suitable control DNA sample for my CUT&RUN NG-seq experiment?