Technical support
Results (652)
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 species reactivity of the SUMO remnant PTMScan® kits that utilize WaLP for peptide digest?
Wild type alpha-lytic protease (WaLP) is a serine endopeptidase that cleaves at the carboxyl terminal side of amino acids alanine, serine, threonine, and valine.
WaLP recognizes and cleaves SUM...
What is your protocol for PNGase F treatment?
To deglycosylate a protein of interest in native conditions.
Reagents:
PNGase F, NEB P0704L (https://...
Are the Protein A Magnetic Beads #73778 and the Protein G Magnetic Beads #70024 hydrophilic or hydrophobic?
The base particle of the Protein A Magnetic Beads #73778 and the Protein G Magnetic Beads #70024 is somewhat hydrophilic. It is difficult to predict the hydrophilicity or hydrophobicity of the protein...
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...
How many reads per cell should I allocate for the InTraSeq™ library?
The number of reads for the InTraSeq protein library should be at least 5,000 reads per cell.
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...
Should I be concerned that my Concanavalin A beads "clump together" in some of my CUT&RUN experiments?
The Concanavalin A beads can clump together rather easily, especially when the cells become lysed. To avoid this, make sure your cells are healthy and be sure to treat them very gently during the cell...
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 ...
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...
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...
Is the VersicanG1-Fc (Rabbit IgG1) #15933 product homologous with all species?
The VersicanG1-Fc (Rabbit IgG1) #15933 contained in the Hyaluronan Complete Tissue Staining Kit (HRP) #38822 and Hyaluronan Complete Tissue Staining Kit (Alexa Fluor® 488) #53721 has been tested succe...
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 ...
Which cleaved caspase-3 (Asp175) antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with mouse samples?
We have several antibodies for detecting caspase-3 when cleaved at Asp175 that are validated for IHC-P with mouse samples. These are:
- Cleaved Caspase-3 (Asp175) Antibody #9661
- Cleaved ...
Can the Senescence β-Galactosidase Staining Kit #9860 be used with frozen tissue?
While we have not tested the Senescence β-Galactosidase Staining Kit #9860 with frozen tissue in-house, we have spoken with numerous customers who have done so with success using our recommended cell ...
Which cleaved caspase-3 (Asp175) antibody do you suggest for paraffin-embedded immunohistochemistry (IHC-P) with human samples?
We have several antibodies for detecting caspase-3 when cleaved at Asp175 that are validated for IHC-P with human samples. These are:
- Cleaved Caspase-3 (Asp175) Antibody #9661
- Cleaved ...
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 RRIDs (Research Resource Identifiers) can be found in the RRID Portal (https://scicrunch.org/resources). Simply search for the product number, clone number, or target along with 'Cell Signaling T... 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... The choice of AMPKα total antibody depends on the subunit of interest and the secondary antibody to be used. Cell Signaling Technology (CST) has five AMPKα antibodies that detect total protein:
Where can I find the RRID numbers for CST antibodies?
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?
Which total AMPKα antibody do you suggest for my western blot experiment?
<...
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 positive and negative controls can I use to demonstrate that my CUT&Tag experiment is successful?
We recommend using a CUT&Tag-validated positive control antibody, like our Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb #9751, in your experiment to confirm that your CUT&Tag assay is worki...
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...
Which isoforms does the ba1/AIF-1 (E4O4W) XP® Rabbit mAb #17198 detect?
Product Iba1/AIF-1 (E4O4W) XP® Rabbit mAb #17198 is predicted to detect isoform 1 and isoform 2 of the Ionized calcium-binding adaptor molecule 1 (Iba1), also known as allograft inflammator...
Will your GFP (D5.1) Rabbit mAb #2956 detect EGFP, eBFP2, and/or mCerulean3?
Our GFP (D5.1) Rabbit mAb #2956 is expected to detect EGFP, eBFP2, and mCerulean3.
Should digitonin be included in my wash buffers for my entire CUT&Tag experiment?
Yes, digitonin needs to be present in all wash buffers during the entire CUT&Tag experiment. This is because cell membrane permeabilization caused by digitonin is reversible. Removing or lowering ...
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 #4691
- Akt (pan)...
Which cleaved caspase-3 (Asp175) antibody do you suggest for immunofluorescent immunocytochemistry (IF-IC) with human samples?
We have several antibodies for detecting Cleaved Caspase-3 (Asp175) that are validated for IF-IC with human samples. These are:
- Cleaved Caspase-3 (Asp175) Antibody #9661
- Cleaved Caspas...
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.
Does CUT&RUN show a bias toward euchromatin or heterochromatin?
We do not observe any bias toward euchromatin or heterochromatin in our assays. In CUT&RUN, the target-specific antibody recruits the pAG-MNase and directs digestion to chromatin that is directly ...