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Concanavalin A beads can clump together rather easily, especially when cells become lysed. Make sure your cells are healthy and be sure to treat them very gently during the cell wash. We also recommend limiting the room temperature incubation time of the cell suspension with Concanavalin A beads to 5 minutes. Resting the tubes at the appropriate incubation temperatures in lieu of rotating or rocking the tubes during the incubations for cell-bead binding, antibody binding, and pAG-Tn5 enzyme binding helps to mitigate bead clumping and does not have any adverse effects on sequencing depth, identified peak numbers, or binding patterns.With all that being said, our internal testing has shown that bead clumping does not negatively affect the final results of CUT&Tag experiments.
The privacy policy currently in place for California residents from Cell Signaling Technology, Inc.
随着流式细胞术近几年的飞速发展,它已成为生物科学领域最有力的分析技术之一。对于这一技术,有的人却望尘莫及,问题层出不穷,比如,胞内染色染不出,找不到合适的抗体,修饰蛋白不知道怎么刺激等等。其实,跳出墨守陈规的流式应用,深度挖掘更深层次的流式实验,助力科研及临床的发展,是所有 Flower 为之奋斗的目标。本讲座将带您了解 Cell Signaling Technology (CST) 流式产品在细胞通路研究中的独到之处,让我们一起在广阔的流式世界中,携手同行,共辟蹊径
Multiplex IHC - Being able to visualize multiple targets simultaneously is important in research fields like immunology & oncology. Learn more here.
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 remaining peptides for repeat LC-MS/MS runs, if necessary.
Need to choose fluorophores and antibodies when designing your immunofluorescence experiment? Our immunofluorescence expert has some advice.
How can intracellular and extracellular protein readouts be combined in flow cytometry? This video offers protocol advice from our flow expert.
Yes. We have shown that our CUT&RUN Assay Kit #86652 works with mouse liver, mouse brain, and mouse heart tissues with as little as 1mg of tissue for each CUT&RUN reaction. Fresh tissue samples work with histone modifications and high abundance DNA binding factors like CTCF. However, for optimal results with transcription factors, cofactors, and difficult tissue types, a light to medium fixation (0.1% formaldehyde, 2-10 min) should be performed and slightly more starting material (2.5mg-5mg) should be used. Fixed tissues can be frozen for up to 6 months before use in the CUT&RUN assay.
"Mouse on mouse" (MOM) background is caused by the anti-mouse secondary antibody binding to endogenous mouse IgG expressed in the tissue resulting in non-specific staining, which can complicate the interpretation of your results. A secondary-only control should be included to assess the potential amount of MOM background in your samples. There are commercially available kits and blocking solutions available that are designed to enable the use of mouse antibodies on mouse tissue. These include M.O.M.® (Mouse on Mouse) Blocking Reagent from Vector Labs and UltraVision™ Quanto Mouse on Mouse from Thermo. We do not routinely use these products in-house so we are not able to recommend one over another.
Yes, our CaMKII (pan) (D11A10) Rabbit mAb #4436 detects the delta isoform.
Yes, you can use BSA #9998 as a carrier for cytokines. It should be dissolved into 1X PBS and sterile filtered.
Cell viability assays measure the population of live, viable cells within a sample. Typically, viability assays measure markers of cell health, including cellular metabolism, ATP levels, and cell proliferation.
We have had customers perform this assay using frozen cell pellets, prior to adding any fractionation buffer, and the subsequent fractionation worked just fine. Once you start fractionating, we recommend following the protocol through to the end. We do not recommend leaving the cell pellets frozen for a prolonged period of time because, even frozen, proteases and phosphatases can slowly degrade the sample. However, leaving the frozen pellet over a weekend or a few days should be ok. Results may vary depending on the stability of your specific targets of interest.
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 other assays following SignalStar, such as hematoxylin and eosin (H&E) staining.
Yes. In our protocol, we recommend a probe-based sonicator because it is less expensive and widely used in many labs. However, you can use a Covaris water bath sonicator. You will still need to optimize the sonication program based on your DNA image, since you want to avoid over-sonicating when performing sonication-ChIP as this can damage chromatin quality/integrity. When looking at sonication results, chromatin DNA fragments should appear as a smear typically ranging between 200 to 1000 bp in size. We recommend you use the condition that requires the minimal sonication cycles, as long as the majority of DNA is less than 1000bp.
作为指导方针,本次讲座将与您分享CST流式细胞术专家在日常的试验工作中经常使用的经验和技巧。
Why are antigen retrieval and antibody diluent important for the immunohistochemistry protocol? Our IHC expert offers protocol advice.
Regrettably, we cannot offer any firm guarantees when glycerol formulated antibodies have been frozen. However, we have performed stability testing by western blot for a representative group of rabbit polyclonal, rabbit monoclonal, and mouse monoclonal antibodies, and found that two freeze/thaw cycles on dry ice (-78.5C) did not affect their activity.
The antigens for our COX IV Antibody #4844 and COX IV (3E11) Rabbit mAb #4850 are 100% conserved with COX IV isoform 1. There is less than 20% homology of these antigens with COX IV isoform 2, therefore we do not expect these antibodies to detect isoform 2.The antigen for our COX IV (4D11-B3-E8) Mouse mAb #11967 is also 100% conserved with COX IV isoform 1. However, the antigen used to produce #11967 is 79% homologous with COX IV isoform 2. Therefore there may be cross-reactivity of this antibody with isoform 2, but we have not tested this in-house and cannot guarantee it.
本讲座将为大家系统地介绍cGAS-STING信号通路在天然免疫和肿瘤免疫研究领域的广泛作用,以及作为干预靶标的巨大潜力。
Apoptosis is a type of programmed cell death that occurs normally throughout the lifespan and can also occur in response to harmful stimuli. Apoptotic cells are identified by their altered morphology, caspase activation, and the presence of damaged DNA.
本讲座全面讲解如何深入开展信号通路的研究,从大数据挖掘到锁定目标信号通路,进一步探讨疾病分子机制,并涉及到相关主流的实验技术,如修饰蛋白质组数据库使用、WB、ChIP、IHC、PTMScan蛋白质翻译后修饰高通量筛选技术等。
Our Insulin (C27C9) Rabbit mAb #3014 was produced with an antigenic peptide corresponding to residues within the insulin B chain of the human insulin protein (UniProt ID #P01308; https://www.uniprot.org/uniprot/P01308). Therefore, it is expected to recognize both proinsulin and mature insulin. While this antibody is not approved for use in western blotting, it should be able to detect the proinsulin protein if the gel is run carefully enough.We also offer an Insulin (L6B10) Mouse mAb #8138, which is approved for use in western blotting on rat samples. This antibody detects both proinsulin and insulin as well.
PTMScan Discovery uses immunoaffinity LC-MS with motif antibodies to identify and quantify PTMs on cellular proteins.
本讲座将概述细胞代谢相关的信号通路,并举例说明系列重要细胞代谢研究靶点,和精彩研究案例思路
Yes, you may freeze your cells and pool them for a later CUT&RUN experiment. However, we would suggest a light fixation using 0.1% formaldehyde for 2 min before freezing the cells, in case the protein-DNA association inside the nuclei changes during the freeze-thaw of the cells.
Marketing department will respond to signal pathway request. Link to tech support.
细胞焦亡与其他多种细胞程序性死亡形式之间存在相互联系和转化,共同构成细胞程序性死亡的复杂体系,维持细胞和机体的内稳态。
When developing the Malachite Green Phosphate Detection Kit #12776, we successfully used the Millipore Centrifuge filters AMICON ULTRA 0.5mL, 10,000 NMWL, product # UFC501096.