Since the paratopes of the antibody fragments are identical to that of the corresponding full-length monoclonal antibodies, their specificity for scFv linkers remains unchang...
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 chromat...
Which phospho-AMPKα antibody do you suggest for my western blot experiment?
The selection of a phospho-AMPKα antibody depends on the subunit of interest. Cell Signaling Technology has four antibodies that target phospho-AMPKα:
Why is a fluorescent conjugate of a rabbit antibody giving no signal on compensation beads in flow cytometry?
If you are using compensation beads in your flow cytometry experiment and observe no signal with one of our fluorescent conjugates of a rabbit antibody, this may be due to the reac...
Is prewashing the Protein A Magnetic Beads #73778 and Protein G Magnetic Beads #70024 necessary for my IP experiment?
We recommend washing the Protein A and Protein G magnetic beads prior to use to help clear them of any potential increase in background due to the buffer in which they are provided...
What isoforms does the TGF-β Antibody #3711 detect and will it detect the 25kDa mature form of TGF-β?
The TGF-β Antibody #3711 detects recombinant TGF-β1, TGF-β2, and TGF-β3. The antibody also detects endogenous levels of the TGF-β1 precursor proteins. The different sizes we have l...
How were the sequences of the PTMScan® Control peptides chosen?
The sequences of the synthetic peptides were based on modified peptides that we have consistently recovered and identified in human and mouse sam...
Is TMT labeling compatible with PTMScan®?
We understand some customers choose to pursue TMT labeling for PTMScan proteomics analysis, but we do not have a defined protocol for this method. However, there are two questi...
Why isn't my bottle of Cell Lysis Buffer (10X) #9803 frozen?
Occasionally, Cell Lysis Buffer (10X) #9803 stored at -20C may not appear frozen, which is not a reason for concern. This phenomenon is known as nucleation, or the initial formatio...
Why was the Caspase-3 (8G10) Rabbit mAb #9665 discontinued?
Additional internal testing has shown that the Caspase-3 (8G10) Rabbit mAb #9665 recognizes both Caspase-3 and Caspase-7. When this antibody was released to the market, we were una...
Why are the beads from PTMScan® HS Kits aggregating against the sides of the tube?
Through in-house testing, we have found that the magnetic beads from our PTMScan HS kits aggregate and stick to the walls of some specific brands of tubes. Moreover, they cannot be...
How much sample is required for a PTMScan® experiment?
We recommend using 10 mg of protein per immunoaffinity purification (IAP) sample for our standard PTMScan kits. For the PTMScan HS product line, 1 mg of starting material is optima...
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:
What is your protocol for treating western blot membranes with CIP?
CIP-treated membranes are prepared with the following protocol:
REAGENTS:
Quick CIP - #M0525 (New England Biolabs)
rCutSmart™ Buffer - #B6004 (New England Biolabs)...
Where can I find the recommended primary antibody dilution buffer for western blotting (BSA or milk)?
The recommended primary antibody dilution buffer for western blotting (i.e., BSA or nonfat dry milk) can be found either in a grey box on the bottom of the first page of the produc...
What can I use as a loading control with the Cell Fractionation Kit #9038?
Due to the nature of the Cell Fractionation Kit #9038 and the localization of each of the proteins in the Loading Control Sampler Kit #5142, the proteins will not be equally expres...
How is the PTMScan® HS Ubiquitin/SUMO Remnant Motif (K-ε-GG) Kit #59322 different from traditional, agarose bead PTMScan®?
The PTMScan® HS Ubiquitin/SUMO Remnant Motif (K-ε-GG) Kit #59322 uses the same antibody as in the traditional PTMScan Ubiquitin Remnant Motif (K-ε-GG) Kit #5562 and PTMScan Pilot U...
Does the CST CUT&Tag kit work with isolated nuclei?
Our CUT&Tag kit is optimized for use with whole cells and works with a large number of cell lines, both adherent and suspension. While our kit works with pre-isolated nuclei be...
What is the pH of SignalStain® Antibody Diluent #8112?
The pH of our SignalStain® Antibody Diluent #8112 has a range between 7.38-7.45.
Are the light chains of the β-Actin (8H10D10) Mouse mAb #3700 kappa or lambda?
The β-Actin (8H10D10) Mouse mAb #3700 has kappa light chains.
Why do I see a doublet when using my total Histone H3 antibody for western blot?
We see this doublet phenomenon often in a number of different cell lines that we test with our Histone H3 antibodies. It does appear to be sample dependent. The doublet may be due ...
What is the expected CUT&RUN DNA yield when starting with extremely low cell numbers, such as 5,000 - 10,000 cells?
If you start with 100,000 cells, you typically get 0.6-6ng DNA. However, if you start with 5,000 - 10,000 cells, the DNA yield can be as low as 0.1-0.2 ng or even undetectable by t...
What are the lower and higher limits of detection for your ELISA kits?
Our in-house threshold for a positive signal is an OD reading of 1.5. Each ELISA kit has a different lower and higher limit of signal detection, based on the sensitivity profile of...
How many LC-MS/MS injections are obtained from a PTMScan® enrichment?
Each PTMScan® enrichment contains sufficient peptides for three to four LC-MS/MS runs. Therefore, one enrichment can produce enough peptides for technical replicates and enough rem...
