Technical support
Results (655)
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...
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...
What is your protocol for PNGase F treatment?
To deglycosylate a protein of interest in native conditions.
Reagents:
PNGase F, NEB P0704L (https://...
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 ...
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...
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 ...
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...
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.
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...
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 possible to perform ... 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?
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 because the emission spe...
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...
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 isotopes has no effect ...
How can I place an order using a quote?
For our US-based customers, quotes may be used on orders placed by phone at 877-616-2355 or by email at [email protected].
When...
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. These are p44/42 MA...
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 fractionating, we recommen...
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 scientists that norma...
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 compared to the 1:250 ...
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...
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...
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....
How do I determine the concentration of my fractionated samples using the Cell Fractionation Kit #9038?
Since the buffers in the Cell Fractionation Kit #9038 contain SDS, you should use a commercially available ready-to-use, detergent- and reducing agent-compatible assay to quickly measure total protein...
Why is there an additional band at 24kDa using the Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb #9718?
The extra band(s) around 24 kDa using our Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb #9718 are most likely mono-ubiquitylated H2AX. H2AX, like H2A, is mono-ubiquitylated.
What is the binding capacity of the antibody-bead conjugates for the agarose PTMScan® kits?
The background binding of non-PTM peptides is typically 50% or more of the total peptides bound to the antibody beads in our agarose PTMScan® kits. This has been improved to less than 50% background o...
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...
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 ...