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Our Phospho-Akt (Thr308) (C31E5E) Rabbit mAb #2965 detects over-expressed levels of PKCalpha and beta. It also detects endogenous levels of PKC in Jurkat + CalA extract and possibly weakly in other endogenous models tested. Our Phospho-Akt (Thr308) (L32A4) Mouse mAb #5106 and Phospho-Akt (Thr308) Antibody #9275 detect over-expressed levels of PKCalpha and beta, but do not detect endogenous PKC. Our Phospho-Akt (Thr308) (244F9) Rabbit mAb #4056 detects over-expressed levels of PKCalpha and beta and may also weakly detect endogenous PKC in Jurkat + CalA extract, but not in other endogenous models tested. Our Phospho-Akt (Thr308) (D25E6) XP Rabbit mAb #13038 does not detect over-expressed PKC and is the best choice of the Phospho-Akt (Thr308) antibodies if this is a concern.
This video presents tips to help your immunoprecipitation experiment: how to avoid IgG masking; optimizing epitope tags; experimental design (controls); what to consider when choosing lysis buffers, antibodies, and beads.
This video describes the simplified PTMScan® HS workflow for LC-MS analysis of ubiquitination and SUMOylation that improves readout sensitivity, reduces sample prep time, and eliminates lyophilization, 9M urea, and antibody elution steps.
SignalStar™ Multiplex IHC Kits & Reagents have not yet been validated for use in frozen tissues. We are currently in the process of validating our antibodies and protocols for use in fresh or frozen tissue.
Multimers are likely to demonstrate higher apparent affinity due to avidity effects. However, an exact recommendation for a competition assay would depend on other assay parameters, specifically assay format (in-tandem, classical sandwich, or premix), tags present on other proteins being used, and detection method.If you want to immobilize ACE2, we recommend using the Human ACE2 (18-652) Recombinant Protein (mFc-Tag) #83986 due to the ease of immobilization by protein A or anti-Fc. If you want ACE2 in solution, we recommend using the Human ACE2 (multimeric) (18-652) Recombinant Protein #85054 if you are immobilizing other proteins by protein A or anti-Fc. This is because the multimeric protein does not cross-react at protein A or anti-Fc surfaces. If you would like ACE2 in solution but are not using protein A or anti-Fc surfaces for immobilization, either protein should work well. Finally, if you want the most potential avidity in solution, then the multimeric protein is preferable.
What to know about widefield versus confocal imaging settings for your microscope to get optimal immunofluorescence data.
The concentration of the pAG-MNase Enzyme #57813 in our CUT&RUN kits is 25ug/ml.
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.
Learn about the power and applications of recombinant antibodies in research. Click here to find out more and revolutionize your experiments.
It should not matter whether you spike in IgG2a or IgG1 antibodies when using the SARS-CoV-2 Spike RBD-ACE2 Blocking Antibody Detection ELISA Kit #69486, as the kit is measuring the ability of the sample to block the ACE2-RBD interaction and is not directly measuring the antibodies. Specifically, this kit is designed to detect blocking antibodies present in human serum or plasma samples.
Whether you're learning a new technique for the first time, or need to troubleshoot problems in your experiments, CST scientists are here to provide guidance. The same application experts that develop and validate our antibodies are here to share their k
The predicted molecular weight of PD-L2 is 31 kDa (based on amino acid composition). However, in most models, PD-L2 migrates at an observed molecular weight of 45-60 kDa on Tris-Glycine gels. This discrepancy between the predicted and the observed molecular weights is a result of glycosylation [see Wang, H et al. (2017) Oncoimmunology 6(7), e1327494 (PMID: 28811964; https://www.ncbi.nlm.nih.gov/pubmed/28811964)].
How does protein expression level factor in your ability to detect proteins and post-translational modifications by western blot?
Each buffer in the Cell Fractionation Kit #9038 contains detergents to help isolate different cellular fractions. The Cytoplasmic Isolation Buffer (CIB) contains a non-ionic detergent used as a cell permeability agent, which causes the plasma membrane to become permeable enough to release cytosolic proteins but also leave other organelles intact. The Membrane Isolation Buffer (MIB) contains another non-ionic detergent used to lyse and solubilize membrane proteins. Finally, the Cytoskeletal/Nuclear Isolation Buffer (CyNIB) contains a denaturing detergent. The specificity of each buffer allows the user to generate strong, clean fractions.
Our COX IV (3E11) Rabbit mAb #4850 will specifically label human cells and not mouse, and our COX IV (D6I4K) Rabbit mAb (Rodent Specific) #38563 will specifically label mouse cells and not human.
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.
We use the Octet® RED96.
We typically recommend the PathScan® Sandwich ELISA Lysis Buffer (1X) #7018 that comes with the Glucose-6-Phosphate Dehydrogenase (G6PD) Activity Assay Kit #12581 as it has been validated for use with the kit. The PathScan® Sandwich ELISA Lysis Buffer (1X) #7018 was made specifically for this assay kit and for ELISA formats. We have not tested any other lysis buffers with the Glucose-6-Phosphate Dehydrogenase (G6PD) Activity Assay Kit #12581, so we cannot guarantee the performance. The ingredients of the PathScan® Sandwich ELISA Lysis Buffer (1X) #7018 are as follows: 20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM Na2 EDTA, 1 mM EGTA, 1% Triton, 20 mM sodium Pyrophosphate, 25 mM Sodium Fluoride, 1 mM β-glycerophosphate, 1 mM Na3VO4, 1 ug/ml leupeptin.
BrdU Cell Proliferation Chemiluminescent Protocol: easy to follow directions describing the step by step experimental procedure.
Immunohistochemistry Protocol (Paraffin): easy to follow directions describing the step by step experimental procedure.
The solution formulation of the PTMScan Control peptides is 25% acetonitrile, 74.9% water, 0.1% trifluoroacetic acid (V:V).
Fluorescent Western Immunoblotting Protocol (Primary Ab Incubation In BSA): easy to follow directions describing the step by step experimental procedure.
A novel study from the laboratory of Daniel Figeys begins to unravel the complexity of the microbiome of pediatric patients with Crohn’s disease.
Protocol for Combined Staining of Intracellular Proteins and Cell Surface Markers in Blood: easy to follow directions describing the step by step experimental procedure.
Flow Cytometry Protocol: easy to follow directions describing the step by step experimental procedure.
Not all cancer cells are equal, they evolve in response to selective pressure driven by accumulation of mutations.
Caspase-3 Activity Protocol: easy to follow directions describing the step by step experimental procedure.
In this webinar, the speakers will explore how tumors exploit immune modulatory mechanisms to generate and thrive in their own immunosuppressive microenvironment. In addition, they will:
When performing immunoprecipitation (IP) followed by western blotting, the denatured IgG light and heavy chains of the primary antibody used for IP run at approximately 25 and 50 kD, respectively, on the subsequent western blot and can often obscure bands of proteins that have similar molecular weights. The use of antibodies from the same animal source for both the IP and the subsequent western blot is the cause of this result since the secondary antibody used in the western blot not only detects the native IgG used for the primary antibody incubation, but also detects the denatured heavy and light chains of the antibody used for the IP. There are several methods to avoid this outcome (ranked from most to least reliable):
Yes, the Fc-Tag should be able to be purified with Protein A or G beads.