Technical Support Articles
Results (709)
Do you have mouse-reactive antibodies available for SignalStar™ Multiplex IHC?
Yes, please select “Mouse” in the first step of the SignalStar Multiplex IH...
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...
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 ...
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 ...
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 & Reagents have not been validated for use in combination with other assays. The SignalStar technology does not destroy the tissue, so it may be ... 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 ... 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:
Can my slides be used for other assays after SignalStar™ Multiplex IHC?
Can I autoclave the 6-Tube Magnetic Separation Rack #7017?
Which phospho-Akt (Ser473) antibody do you suggest for Immunofluorescence (Immunocytochemistry) (IF-IC) with human samples?
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...
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 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 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 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...
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...
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 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...
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...
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...
Does CST have a protocol for treating cells with poly(dA:dT)?
The suggested protocol for treating cells with poly(dA:dT) can be found below:
Poly(dA:dT) Treatment Protocol (5 μg/mL final concentra...
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...
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...
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...
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...
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 ...
Does the CUT&Tag assay show a bias toward euchromatin or heterochromatin?
With the appropriate salt concentration in the tagmention buffer, we don't observe bias toward euchromatin or heterochromatin in our assays. In CUT&Tag, the target-specific...
What is the composition of the DNA Elution Buffer #10009?
The composition of our DNA Elution Buffer #10009 is 10 mM Tris-Cl, pH 8.5.
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 Ribo...
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, on...
How do I avoid batch effects and what sample run order should I use for PTMScan® experiments?
To avoid intra-sample batch effects during liquid chromatography with tandem mass spectrometry (LC-MS/MS), we recommend a run order that involves running one technical replicate of...
Will your Anti-biotin, HRP-linked Antibody #7075 cross-react with endogenous biotin in cell lysates?
Our Anti-biotin, HRP-linked Antibody #7075 is intended for use with our Biotinylated Protein Ladder Detection Pack #7727. However, it is possible for this antibody to also detec...