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All of our siRNAs are single duplexes. We discontinued pools a number of years ago to avoid off-target effects.
It is critical that our Phospho-p53 (Ser15) Antibody #9284 be diluted into 5% w/v non-fat dry milk, 1X TBS, 0.1% Tween® 20 and incubated at 4°C with gentle shaking, overnight. Diluting the antibody in BSA will result in high background on the blot.
Cell Lysis Buffer (10X) #9803 will interfere with the Bradford protein assay. This is because it contains detergent. To make the assay somewhat compatible, dilute your protein quantitation samples at least 10-fold with water or PBS to reduce the amount of detergent and consequentially lower the interference. This ten-fold dilution may still yield background absorbance. We recommend employing an alternative protein assay that is compatible with #9803, such as the BCA Protein Assay Kit #7780.
PEG (polyethylene glycol) has been added to our Anti-rabbit IgG (H+L) (DyLight™ 800 4X PEG Conjugate) #5151 and Anti-mouse IgG (H+L) (DyLight™ 800 4X PEG Conjugate) #5257 to increase their solubility and stability for various applications, including cellular and in vivo imaging. Internal Fluorescent Western and In-cell Western testing shows that the addition of PEG to these secondaries not only significantly increases signal but also decreases background fluorescence.
There are 3 isoforms of Estrogen Receptor α (ER66, ER46, and ER36). Our Estrogen Receptor α (D8H8) Rabbit mAb #8644 will recognize ER66 and ER46, but it will not recognize ER36.
It is possible to see a doublet at ~100 kDa for Grp94 due to post-translational modifications. Here is a link to our PhosphoSitePlus database detailing the possible post-translational modifications for Grp94: https://www.phosphosite.org/proteinAction.action?id=6928&showAllSites=true.
HPSE is synthesized as a 65 kDa inactive precursor that undergoes proteolytic processing, yielding 8 kDa and 50 kDa protein subunits that heterodimerize to form an active enzyme. The 65kDa proform and the 50 kDa subunit (amino acids 158-543) are detected by the HPSE (E3N3H) Rabbit mAb #99756, while the 8kDa subunit (amino acids 36-109) is not.
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.
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)].
We have several antibodies for β-actin that are validated for IHC-P in human samples. These are β-Actin (13E5) Rabbit mAb #4970 and β-Actin (8H10D10) Mouse mAb #3700. Both work well and there is no preference for this application.
The recombinant protein utilized by the SARS-CoV-2 Spike Protein Serological IgG ELISA Kit #20154 consists of the complete ectodomain, including all of S1 and S2 of the spike protein. It is not the full-length spike protein but rather the complete ectodomain, which is the domain of a membrane protein that extends into the extracellular space. The ectodomain is usually the part of a protein that initiates contact with surfaces and leads to signal transduction. In SARS-CoV-2, the ectodomain of the spike protein is responsible for attachment to and entry into cells during infection.
Glucose-6-phosphate (G6P), in the presence of NADP, is oxidized by Glucose-6-Phosphate Dehydrogenase (G6PD) to generate 6-phosphogluconolactone and NADPH. The NADPH is then amplified by the diaphorase-cycling system to produce highly fluorescent resorufin molecules. We assume that by measuring the resulting NADPH, the signal is proportional to the G6PD activity in your sample. 6-Phosphogluconate dehydrogenase (6PGD) also uses NADP+ to generate NADPH, and therefore, may contribute to the overall signal. As stated in the Specificity / Sensitivity section on the product webpage, the Glucose-6-Phosphate Dehydrogenase (G6PD) Activity Assay Kit #12581 detects sample G6PD activity, however the presence of NADH and NADPH may interfere with the assay. Therefore, it may be difficult to distinguish between G6PD and 6PGD activity when using this kit.
We recommend that you complete the BrdU cell proliferation assay as soon as you have the time. However, if you are unable to complete the experiment during the recommended time, you can complete the experiment through Step B: BrdU Incorporation and then pause. To do this, please follow the steps below:1. Add the fixing/denaturing solution and incubate for 30 minutes (Part C: BrdU Assay Step 1).2. Remove your solution and dry your plate at room temperature for 30 minutes. 3. Cover plates with parafilm and store at 4C.It is important to make sure your plate is completely dry before you cover with the parafilm and store it. Dry plates can be stored for up to 1 month at 4C, however, the signal of the assay may decrease slightly.
We have several antibodies for cleaved PARP that are validated for IHC-P in human samples. These are Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb #5625 and Cleaved-PARP (Asp214) (E2T4K) Mouse mAb #32563. Both work well and there is no preference for this application.
The main difference between our SimpleChIP Enzymatic Chromatin IP Kits (#9002 & #9003) and our SimpleChIP Plus Enzymatic Chromatin IP Kits (#9004 & #9005) is that the SimpleChIP Plus Enzymatic Chromatin IP Kits contain a protocol for fixing and preparing chromatin from tissues and the required additional material to do so (which is the reason for the price difference).
Discover the TGF Beta signaling pathway and impact on cell growth & tissue homeostasis. Learn here the mechanisms behind this vital signaling cascade.
The YAP/TAZ (D24E4) Rabbit mAb #8418 antibody is generated using the TAZ human protein sequence (UniProt ID - Q9GZV5). However, the YAP sequence (UniProt ID - P46937) in the same region is only approximately 77% identical to the TAZ sequence used, which may affect how well the antibody binds to YAP.
ChIP-seq is a powerful technique that combines ChIP and next-generation sequencing (NGS) to study DNA-protein interactions across the entire genome, but it is only as good as the quality of the antibody used in the ChIP experiment.