Basket Updated
0
Items added
Learn about the antibodies, custom reagents, and ready-to-use cell assay kits to support your CAR-T therapy development.
The calculated molar extinction coefficient of rabbit IgG at 280 nm is approximately 210,000 M-1cm-1.
Products Terms and Conditions, Except for Bioconjugation - Cell Signaling Technology
Bioconjugation Services Terms and Conditions - Cell Signaling Technology
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 listed represent the precursor TGF-β protein (45kDa), the mature dimer form (25kDa) and the monomer (12kDa).We do not usually detect the mature TGF-β at 25 kDa because mature TGF-β is composed of two polypeptides that are disulfide-linked. Therefore under reducing conditions, these polypeptides are separated and observed as a single band at 12 kDa. However, we have shown that the TGF-β Antibody #3711 can detect the mature form of TGF-β by using recombinant protein.TGF-β is rapidly exported from the cell and therefore may be difficult to detect in tissue or cell lysates. In human platelet samples, we observe two strong bands, with the larger band likely being the precursor (uncleaved) form at 45kDa and the smaller being the monomer at 12kDa.
In addition to product development and production, CST is also actively engaged in the development of new technologies for signaling analysis as well as mechanistic cell biology research.
Our Ubiquitin (P4D1) Mouse mAb #3936 weakly detects free Ubiquitin. The Ubiquitin signal can be increased by using a semi-dry-transfer and autoclaving (30 min. wet cycle) or boiling the membrane for 30 minutes after transfer.
In endogenous samples, the PINK1 precursor protein (~60kDa), is imported into mitochondria where it is processed into a cleaved form (~50kDa) that is transported back to the cytoplasm to be rapidly degraded by the proteasome. Since PINK1 is degraded so quickly, detecting endogenous PINK1 in untreated cell extracts is unlikely. In order to observe PINK1 in endogenous samples, we use the induction of mitophagy by treatment with CCCP, which leads to the accumulation of cleaved PINK1. However, we do observe two bands, representing both the precursor and cleaved forms, when using samples transfected with full-length PINK1. This is due to the high levels of PINK1 in these samples.
Video: the updated CUT&RUN kit provides improved results, confirmed compatibility with fixed frozen cells, tissues, and primary cells, uses even fewer cells.
The background or number of non-PTM peptides in each enrichment is primarily due to non-specific interactions with the beads the antibody is conjugated to. Therefore, this background is also non-depletable by methods such as pre-clearing.The level of background peptides can range considerably depending on the PTM and the biological source. PTMs that are typically lower in abundance (e.g., phosphotyrosine) can be expected to have a higher background compared to more abundant PTMs like the ubiquitin remnant (or K-e-GG) and acetyl lysine. Peptides derived from cells in culture typically produce more PTM peptides than an equivalent amount of protein from tissue.A major source of background peptides can be carry-over of antibody-beads to the stage tip or small-scale desalting at the end of the protocol. For optimal results, the eluted peptides should be clear of any antibody-beads before loading onto the stage tip. The pH of the peptides in the IAP enrichment step may also affect the specificity…
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 each individual kit. We recommend referring to the lysate titration image on the individual product webpage/datasheet to view our lowest lysate titration and its corresponding OD.
The antigen for the β1-Adrenergic Receptor Antibody #12271 surrounds Gly406 of the human β1-Adrenergic Receptor protein and is contained entirely in the cytoplasmic domain. #12271 detects full-length β1-Adrenergic Receptor as shown on the datasheet (see western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human β1-Adrenergic Receptor (β1AR; +). The smeary bands that can be observed above 50 kD are a result of N-glycosylation. We have not looked into the issue for further details.
Immunoprecipitation Troubleshooting Guide for easy to solve high and low background issues.
Dr. Nathanael Gray of the Stanford Cancer Institute discusses efforts to synthesize and characterize small molecule degraders of cancer targets, including kinases and transcriptional regulators.
How sample type, processing steps, and characteristics of the target protein can impact detection and inform protocol optimization in immunofluorescence.
We have confirmed in-house that our Phospho-p53 (Ser15) Antibody #9284 and Phospho-p53 (Ser15) (D4S1H) Rabbit mAb #12571 cross-react with some prestained protein markers. We are unsure as to why this is occurring. To help avoid this, we recommend using a biotinylated marker without the use of a prestained marker.
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 fragment sizes (i.e. 150, 300, 450, 600 bp). When using antibodies against transcription factors, we see a range of DNA fragment sizes, with the majority being greater than 35 bp.
Privacy
TMT10plex Proteome Analysis in Cell Lines
Phospho-GSK-3β antibodies that detect phosphorylation at Ser9 upon activation in properly treated samples are also capable of detecting the GSK-3β-Tag. Although these antibodies are specific to the Ser9 site of GSK-3β, the GSK-3β-Tag is quite similar to their antigen sequences. CST offers the following phospho-GSK-3β (Ser9) antibodies which also detect the GSK-3β-Tag: Phospho-GSK-3β (Ser9)(D85E12) XP® Rabbit mAb #5558Phospho-GSK-3β (Ser9)(5B3) Rabbit mAb #9323Phospho-GSK-3β (Ser9)(D2Y9Y) Mouse mAb #14630Phospho-GSK-3β (Ser9) Antibody) #9336
All of our siRNAs are single duplexes. We discontinued pools a number of years ago to avoid off-target effects.
What antibodies can be used in a CUT&RUN protocol? What controls should be included to measure signal-to-noise? These and other frequently asked questions are answered in this edition of CST Tech Tips.
We do not routinely test samples prepared from whole tissue. However, customers have successfully used our ELISA kits with their tissue extracts. If you are interested, we can provide a recommended protocol for tissue sample preparation. You will likely need to optimize the amount of tissue that you are loading because of the lower than normal concentration of the target of interest. These kits come with a positive control that can be assayed alongside the tissue sample.
Business Development Request Form
The pathway induction from H202 treatment may be more robust than the LPS treatment. We have previously observed that buffer and temperature conditions at the lysis stage can affect basal levels of PARylation. Notably, lysis at RT yielded lower observable PAR than lysis on ice (4 deg C). Also, performing the initial lysis in a PDE assay buffer (available from several vendors) yielded higher observable PAR than lysis directly into SDS loading buffer with DTT. Lastly, cells overgrown in culture may also show higher basal levels of PAR.
In this video, our experts provide protocol guidance for lysate preparation steps prior to protein electrophoresis and western blotting.
Particulates may occasionally be seen in our Animal-Free Blocking Solution (5X) #15019 due to material becoming caught in the threads of the cap/bottle and drying out over a short period of time. These particulates are of no cause for concern and you may continue to use and dilute the product to the 1X working concentration. You may also heat the product at 37C for 15 minutes to help bring any material back into solution, if required.