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...
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...
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 process of developing cust...
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...
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 ...
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 control, which may som...
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.
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 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 ... 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... 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... 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:
Is PTMScan® compatible with SILAC?
Have species been tested if they are not listed as cross-reactive with a CST antibody on the datasheet?
Which nuclear dyes can be used for In-Cell Western assays in combination with DyLight 680-conjugated secondary antibodies?
Which phospho-Akt (Ser473) antibody do you suggest for Immunofluorescence (Immunocytochemistry) (IF-IC) with human samples?
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 rack with 10% bleach...
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 ...
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 protein:
- I...
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 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...
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...
Why does the TFE3 Antibody #14779 detect multiple bands?
The higher band obtained with the TFE3 Antibody #14779 at approximately 80 kDa represents the sumoylated form of the TFE3 protein, which migrates higher than the unmodified form [see Miller, A....
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...
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 possible to perform ...
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 Ribosomal Protein (5...
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://www.uniprot.org/unipr...
What is the concentration of your Digitonin Solution #16359?
The concentration of our Digitonin Solution #16359 is proprietary. The concentration will vary between lots, as we internally test each lot to match performance before release. This is due to activity...
Do you have human-reactive antibodies available for SignalStar™ Multiplex IHC?
Yes, please select “Human” in the first step of the SignalStar Multiplex IHC Panel Builder ...
Can you provide data that confirms the specificity of your Raptor (24C12) Rabbit mAb #2280?
The specificity of our Raptor (24C12) Rabbit mAb #2280 has been confirmed by siRNA in the following papers:
PMID: 28648777 (https://pubmed.ncbi.nlm.nih.gov/28648777/) Figure 2B
PMID: 249...
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, one can increase the n...
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...
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 Ribosomal Protein (Ser23...
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...