Results Expression and purification of recombinant pINSL3. FLAG-tagged pINSL3 was expressed in silkworm using the BmNPV bacmid system and successfully secreted into larval hemolymph using its native signal peptide (Fig. 1A). Because it was difficult homogenously purify the recombinant protein ...
colorless and transparent. The enriched bands cut on the separation glue after elution were placed in a centrifuge tube and eluted using 3xFlag peptide buffer (150 mM KCl, 10 mM Tris-HCl (pH 8.0), 20% Glycerol, 0.1 mM DTT and 200 μg/ml 3xFlag peptide). The proteins were ...
While we demonstrate that TNIP3 directly interacts with TAK1 and blocks its downstream signaling with its AHD1 domain, whether a TNIP3 AHD1 domain-derived small peptide can recapitulate the function of full-length TNIP3 in this regard deserves to be further investigated. Lastly, whether TNIP3 ...
Furthermore, we required that each protein be detected by at least two unique peptides. Interaction Scoring for FLAG AP-MS For each bait, a cell line was prepared for mass spectrometric analysis as described above. Biological duplicates were grown/treated/processed at different times to maximize ...
The options of “Second peptides,”“Match between runs,” and “Dependent peptides” were enabled. Label-free quantification was used to quantify the difference of protein abundances between different samples [56, 57]. Image processing and statistical analysis Pearson coefficient and line intensity ...
关于产品“ 血蓝蛋白-琼脂糖 Keyhole Limpet Hemocyanin (KLH) (Agarose);CAS:9013-72-3 ”的详情 Name: Keyhole Limpet Hemocyanin (KLH) (Agarose) Desc: Antibodies to small peptide/haptens are gen
(for immunoprecipitation; made in-house), rabbit antibody to p110β peptide (for immunoblot analysis; sc-602; Santa Cruz Biotechnologies), anti-p110γ (4252; Cell Signaling Technologies), anti-p110δ (for immunoprecipitation; EPR386; Abcam), anti-p110δ (for immunoblot analysis; sc-7176) ...
The peptides were analyzed by mass spectrometry and peptide micro-sequencing. Experimental details on the procedures are available upon request. Plasmid constructs and recombinant proteins The pHT-FS1-NTD plasmid was constructed by inserting the NcoI – SmaI fragment from Flag-CSN1 [27] into the ...
peptides with a monomethylated residue at K106 and K200, carrying a mass of + 14.016 Da, respectively.DIn vivo methylation assay of PLK3 using a pan anti-Lys-methyl antibody. Flag-tagged PLK3 WT or mutants were immunoprecipitated with anti-Flag magnetic beads from HEK293T cells co-...
Cleared lysates were then added to anti-Flag M2 Magnetic Beads (Sigma, M8823) followed by end-over-end rotation overnight at 4 °C. Beads were then washed three times in lysis buffer followed by elution in elution buffer (lysis buffer + 0.15 mg ml−1 Flag Peptide (Sigma-...