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β-Actin can be cleaved by caspase-3 between Asp244 and Gly245, which will result in a 32kDa fragment and a 15kDa fragment. Due to the location of its antigen, our β-Actin Antibody #4967 will detect both full-length β-Actin as well as the 32kDa fragment, also known as fractin.
How can intracellular and extracellular protein readouts be combined in flow cytometry? This video offers protocol advice from our flow expert.
The Mouse Anti-Rabbit IgG (Light-Chain Specific) (D4W3E) mAb (HRP Conjugate) #93702 and the Rabbit Anti-Mouse IgG (Light Chain Specific) (D3V2A) mAb (HRP Conjugate) #58802 are specific to kappa light chains, which cover most commercially available monoclonal antibodies. They do not react with lambda light chains.
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.
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.
Why are antigen retrieval and antibody diluent important for the immunohistochemistry protocol? Our IHC expert offers protocol advice.
The GFP Antibody #2555, GFP (4B10) Mouse mAb #2955, GFP (5G4) Mouse mAb #55494, and GFP (D5.1) XP® Rabbit mAb #2956 are not expected to detect Turbo GFP due to low sequence homology.
Yes. Cell fixation is recommended when the cells are very fragile (easily lysed during experiments), when the cellular signaling pathway is sensitive to the Conconavalin A beads used for cell binding, when the target protein binds weakly to DNA, or when you hope to freeze the cell pellets for future use. A light fixation (0.1% formaldehyde for 2 min) is critical for the success of CUT&RUN with fixed cells. A reverse crosslinking step is also required at the end of the assay to remove protein from DNA. We find that live cells generally have better performance than fixed cells in the CUT&RUN assay, therefore, we recommend using live cells if possible, unless fixation is necessary as noted in the above-mentioned scenarios.
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。
本场讲座将逐一介绍多种细胞程序性死亡(凋亡、自噬、程序性坏死、细胞焦亡)相关信号通路关键蛋白和研究策略,并就多种细胞程序性死亡形式之间的联系和转化展开讨论,为大家全面认识细胞程序性死亡和开展系统研究提供参考。
Our Cell Fractionation Kit #9038 provides a fast and efficient way of separating cultured cells into three distinct fractions for analysis by SDS-PAGE and western blotting. We have not tested fractionated samples using mass spectrometry in-house and it is difficult to speculate how well they would work. The buffer contains SDS so you may need to use a commercially available SDS purification kit if you would like to prepare the fractionated samples for mass spectrometry by in-solution digestion; in-gel digestion would account for SDS removal in the extraction already. We cannot guarantee activity by mass spectrometry as it is not the primary intended use of this kit.
该视频重点介绍肿瘤免疫微环境相关的主要参与细胞、调节分子和信号通路,并为各位听众提供在上述领域的主要评价指标和新的研究方法作为参考。
PTMScan Direct uses immunoaffinity LC-MS with a formulation of site specific antibodies to identify and quantify post-translational modifications (PTMs) on cellular proteins in specific pathways.
该视频将重点介绍与细胞骨架、细胞基质动态调控及EMT相关的主要调节分子和信号通路,并为各位听众提供在上述领域的主要评价指标作为参考。
For imaging purposes, we recommend using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb #9664 when working with rodent tissue. We have noted non-specific labeling in specific sub-types of healthy cells in fixed-frozen mouse tissues (e.g. pancreatic alpha-cells) and nuclear background in rat samples with Cleaved Caspase-3 (Asp175) Antibody #9661 and Cleaved Caspase-3 (Asp175) (D3E9) Rabbit mAb #9579.
The ELISA method has many variations, all of which rely on complex of an antigen and an antibody/enzyme conjugate. Signal is generated by turnover of substrate.
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PathScan ELISA kits detect endogenous levels of key signaling proteins using a traditional sandwich-based assay format in phospho-specific or total protein options.
Unfortunately, we cannot confirm that the recombinant SARS-CoV-2 spike proteins we provide are in exactly the same conformation that would be expected in nature, as we do not yet have direct data addressing this issue. That being said, our recombinant SARS-CoV-2 spike proteins are expressed in mammalian cells to try to ensure the presence of secondary (post-translational) modifications, which would not happen in a prokaryotic expression system. Furthermore, we suspect the trimer is closer to native folding than the monomer, but without NMR or X-Ray crystallography, we have not been able to confirm this.
The SARS-CoV-2 Spike RBD-ACE2 Blocking Antibody Detection ELISA Kit #69486 has not been validated for use with any samples except for human serum or plasma. However, it may also be used to assess the blocking activity of non-human antibodies and small molecules. For example, we have had customers successfully use this kit in rat samples. You will likely need to optimize the sample dilution and antibody spike-in concentrations, and we recommend running appropriate controls for each assay (for example, rat sera that should not block activity or concentration-matched, non-relevant antibodies that are spiked in).
Need to choose fluorophores and antibodies when designing your immunofluorescence experiment? Our immunofluorescence expert has some advice.
FastScan ELISA kits detect key signaling proteins using a solution-based assay format and are validated to ensure reproducible results across lots and users.
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
IF-approved antibodies from CST are validated for use with a blocking buffer that contains 5% serum diluted in PBS containing 0.3% Triton X-100. Serum components offer diversity over alternatives such as milk and BSA and may bind and occupy more sources of non-specific protein-protein interaction. Choosing serum from the same species as the host species of your secondary antibody (e.g., goat, donkey) may also help to reduce background originating from endogenous Fc receptors. At CST, we offer fluorochrome-conjugated F(ab’)2 fragment secondary antibodies that reduce the potential for non-specific binding from this source. Therefore, if you are using one of our fluorochrome-conjugated F(ab’)2 fragment secondary antibodies, blocking may not be necessary.
Some common considerations for using the Senescence β-Galactosidase Staining Kit #9860 are as follows:1. Please ensure that the 10X Staining Solution, as well as Supplement A and B, are fully dissolved before diluting it to 1X. The solutions can be heated to 37C for a few minutes to redissolve any precipitate.2. The pH of your final staining solution is very important. A pH of 6.0 is best, but we recommend to keep it between 5.9 and 6.1. A low pH can result in false positives and a high pH can result in false negatives. If necessary, use HCl or NaOH to lower or raise the pH, respectively.3. A dry incubator must be used for X-gal staining as tissue culture incubators have CO2 which can change the pH of the staining solution and affect staining.4. An X-gal solution, when stored at -20C in a polypropylene plastic or glass container and protected from light, is only stable for about one month.
What do your crystals look like under the microscope? Do they look like glass shards?We see these from time to time and they may be undissolved X-Gal which is occurring because there is an abundance of the chemical. It is important to be sure that the original solution is indeed 20mg/ml, as X-Gal can be a bit tricky to weigh out because it is so fluffy. The solutions should also be warmed to 37C prior to diluting, mixing, and adding to cells.Were your plates wrapped in parafilm?This is crucial to help prevent evaporation.Have you checked the age/expiration date of your DMF?DMF tends to expire and this can affect the solubility of the X-Gal.We do have a solution to dissolve the crystals so you may still move forward with your experiment:Create a 50% DMSO and water solution (please use caution when working with DMSO). Remove the staining solution, wash 1X with PBS, and add the DMSO/water solution. Once you observe dissolution of the crystals, remove the DMSO/water solu…
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本次研讨会将通过介绍CST Western Blot标准操作步骤,向您阐释WB操作中的关键点及注意事项,特别是影响最终结果呈现的样本制备和抗体孵育与检测。通过真实失败案例,如无信号、高背景、条带大小不正确、条带弯曲拖尾等,分析WB常见的问题及原因,并提供相应的解决策略。
The top band in the gel image for our Human ACE2 (18-652) Recombinant Protein (mFc-Tag) #83986 likely represents dimerization due to the mFc tag.