coated with anti-stxb antibody 13c4 and using streptavidin-hrp

coated with anti-stxb antibody 13c4 and using streptavidin-hrp

For ELISAs using commercial antibodies, mAbs 3C10 (Toxin Technologies), 13C4 (ATCC Bioresource Center) and Sifin 1 (Sifin) were used as capture antibodies, respectively, and PC1-HRP (Toxin Technologies), which is a combination of Stx1 and Stx2 antibodies conjugated to HRP, was used as the detection antibody. Our most sensitive ELISA, using a ...

3 Children's Hospital, Harvard Medical School, Boston, MA 02115. [email protected] Background: Maintaining DNA integrity by dealkylation is a crucial step to maintain genomic stability. Demethylation by the AlkB dioxygenase represents an important mechanism for repair of N-alkylated nucleotides.

2011-2-1 · Next, the total amount of StxB-sulf2 on the membranes was determined by incubating the membranes with anti-Stx antibody (3C10, Toxin Technology) at 1 µg mL −1 in 2.5% milk for 1 h, followed by 30 min of incubation with a horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG antibody (115-035-003, Jackson Immunoresearch Laboratories, 1 ...

After production, the periplasmic fractions were recovered and tested on Maxisorp microtiter plates coated overnight at 4 °C with BLS-Stx2B or BLS (100 ng/well) and blocked with 3% PBS skimmed milk. Rat monoclonal anti-HA peroxidase conjugated Antibody (recognizes the HA peptide sequence YPYDVPDYA derived from the human influenza virus hemagglutinin protein; Roche) was used for …

2016-4-27 · Selection of anti-Stx2B VHHs. The BLS-Stx2B chimera is a potent immunogen in mice, inducing highly neutralizing anti-Stx2 antibodies 26,27.Thus, we used this immunogen for the development of llama ...

The mouse monoclonal anti-STxB antibody 13C4 was purified from hybridoma cells (ATCC CRL-1794), and the rabbit polyclonal anti-VAMP3 antibody (used at 1:4000 for western blotting) was a gift from Thierry Galli (Institut Jacques Monod, Paris, France). …

开发针对志贺毒素2型(Stx2)的骆驼科单链抗体,具有抗溶血性尿毒症综合征(HUS)的治疗潜力

2017-10-13 · To avoid immune-complex interaction with high-affinity Fc-receptors on macrophages and dendritic cells, a Fab-fragment of the 13C4 antibody was generated that, when prebound to the STxB, also strongly reduced the STxB–Gb 3 interaction on TLC plates (Figure2A) or in the plasma membrane of HeLa cells (our unpublished results).

2019-12-21 · expressed and isolated to high purity, and shown to be efficiently immunodetected using epitope tag antibodies or streptavidin-HRP (Fig. 1B). A well-established phenotype of retrograde transport deficiency on MPR traffic is the redistribution of the

Alexa488 anti-mouse antibody is suitable for 2nd Ab. Hela 細胞 a-adaptin 2011-08-29 評価者へのお問合せ 100/2 WB IF IHC IP SIGMA x200 ― A4325 mouse Mono, IgG2a ...

The quantification of biotinylated STxB was achieved by ELISA on plates coated with anti-STxB antibody 13C4 and using streptavidin-HRP. STxB entry into control cells was highly efficient at 37°C ( ), and was totally abolished at 4°C ( ).

2021-5-25 · U.S. Patent Number 09481726 for Antibodies to TGF-.beta.

2020-2-17 · Horseradish peroxidase (HRP)-tagged antibodies were also purchased from Beckman Coulter and used at a dilution of 1:2,000: donkey anti-rabbit (no. 711035152, lot 136860) and donkey anti …

2014-6-10 · For ELISAs using commercial antibodies, mAbs 3C10 (Toxin Technologies), 13C4 (ATCC Bioresource Center) and Sifin 1 (Sifin) were used as capture antibodies, respectively, and PC1-HRP (Toxin Technologies), which is a combination of Stx1 and Stx2 antibodies conjugated to HRP, was used as the detection antibody. Our most sensitive ELISA, using a ...

The present invention is directed to a method for the synthesis of a bi-functional complex comprising a molecule part and an identifier oligonucleotide part identifying the molecule part. A part of the synthesis method according to the present invention is preferably conducted in one or more organic solvents when a nascent bi-functional complex comprising an optionally protected tag or ...

RES., vol. 146, 1986, pages C1 - C5 NAKADA, H. ET AL.: "Epitopic structure of Tn glycophorin A for an anti-Tn antibody (MLS 128", PROC. NATL. ACAD. SCI. USA, vol. 90, 1993, pages 2495 - 2499 OSINAGA, E. ET AL.: "Analysis of the fine specificity of Tn-binding proteins using synthetic glycopeptide epitopes and a biosensor based on surface plasmon ...

2018-11-3 · The mechanisms underlying the association between immune activation and postpartum depression remained elusive. Although Ω-3 fatty acids possess anti-inflammatory properties, there is limited evidence directly linking the modulating effects of Ω-3 fatty acids on neuroimmune and neurochemistry to the antidepressant actions. A between-groups design was used to assess the …

2019-12-21 · few laboratories imaged antibody uptake to follow retrograde transport from the cell surface to the TGN (e.g., Breusegem and Seaman, 2014), while Johannes and colleagues employed sulfation as a specific modification of the trans-Golgi/TGN to probe Golgi arrival of Shiga toxin B-chain (STxB) tagged with a sulfation motif or of

2007-4-15 · Biotinylated STxB or transferrin was detected using streptavidin-HRP (Roche). The reaction was stopped with 3M sulphuric acid and plate was read at 490 nm. The percentage of internal STxB (or Tf) was determined as the ratio of signal after MESNA reduction (internal ligand) to signal without reduction (total ligand).

2018-9-3 · using Beckman Coulter counter Multisizer 4 with a 30 micron aperture which covers size range from 0.6 micron to 18 micron. Mean particle size of the particles is 3.8 micron with Std. Dev. 2.03. Using the instrument, particle size characterization can be carry out using only 0.5mg of sample with 10 min total measurement time.

2003-4-25 · The anti‐STxB mAb 13C4 and the anti‐Nef mAb MATG 020 were purified as previously described . Anti‐galactosyl‐transferase polyclonal antiserum was provided by E. Berger (Institute of Physiology, University of Zurich, Switzerland) and anti‐Hrs antibodies were kindly provided by S. Urbé (University of Liverpool, UK).

2015-4-16 · Immunoconjugates, bispecific molecules and pharmaceutical compositions comprising the antibodies of the invention are also provided. The invention also provides methods for inhibiting IP-10 activity using the antibodies of the invention, including methods for treating various inflammatory and autoimmune diseases.

After quenching of 2‐mercaptoethanesulfonic acid with iodoacetamide, the cells were lysed in blocking buffer (10 m m Tris/HCl, pH 7.4, 1 m m EDTA, 50 m m NaCl, 0.2% BSA, 0.1% SDS, and 1% Triton X‐100), lysates were transferred into 96‐well plates precoated with mouse monoclonal anti‐STxB 13C4, and STxB–SS–biotin was visualized with ...

2014-4-18 · Shiga toxin-producing Escherichia coli bacteria cause hemorrhagic colitis and hemolytic uremic syndrome in humans. Currently, only supportive treatment is available for diagnosed patients. We show here that 24-h pretreatment with an ether lipid precursor, the alkylglycerol sn-1-O-hexadecylglycerol (HG), protects HEp-2 cells against Shiga toxin and Shiga toxin 2.

2012-8-2 · Shiga toxin (Stx) causes diarrhea-associated hemolytic uremic syndrome by damaging renal microvascular endothelium. The pentameric B subunits of Stx types 1 and 2 (Stx1B and Stx2B) are sufficient to stimulate acute VWF secretion from endothelial cells, but Stx1B and Stx2B exert distinct effects on Ca 2+ and cAMP pathways. Therefore, we investigated other signaling components in StxB …

2019-10-28 · The monoclonal anti-horseradish peroxidase (HRP) antibody was obtained from Stratech Scientific (Luton, United Kingdom). Donkey anti-mouse IgG and donkey anti-rabbit IgG antibodies, conjugated to either fluorescein isothiocyanate (FITC), rhodamine, or …

After washing, excess MESNA was quenched with 150 mM iodoacetamide for 20 min. Cells were lysed in blocking buffer (10 mM Tris [pH 7.4], 1 mM EDTA, 50 mM NaCl, 0.2% BSA, 0.1% SDS, and 1% Triton X-100) before loading on ELISA plates coated either with anti-Tf or anti-STxB antibody (13C4). Biotinylated STxB or transferrin was detected using ...

2014-10-5 · immunolabelled with a mouse monoclonal anti-STxB antibody (13C4) and a cyanin 3-conjugated secondary goat anti-mouse antibody (red), biotin was labelled using FITC-conjugated streptavidin (green) and the Golgi marker Rab6 was immunolabelled using a rabbit monoclonal anti-Rab6 antibody and a cyanin 5-conjugated secondary donkey anti-rabbit antibody (blue). lyophilization, the …

2004-4-1 · Biotinylated STxB or transferrin was detected using streptavidin-HRP (Roche). For the transferrin recycling experiments, biotinylated Tf was internalized into HeLa cells for 40 min at 37°C. After washing on ice, the cells were incubated at 37°C for the indicated times.

2014-4-4 · The monoclonal anti‐STxB antibody 13C4 was purified with Protein‐A‐Sepharose from culture medium of the corresponding mouse hybridoma cells (ATCC). The polyclonal STxB antibody was provided by I. Trowbridge (The Salk Institute) (1:500 wb). Streptavidin–peroxidase (POD) (1:500) was obtained from Roche.

2005-7-1 · Declaration of Financial Interests or Relationships To insure balance, independence, objectivity and scientific rigor in all CME programs it is the policy of the meeting's joint sponsors that any speaker or poster presenter who makes a presentation at a program designated for AMA Physician's Recognition Award (PRA) Category 1 or 2 credit must disclose any real or apparent financial interest …

C9ORF72 depleted cells expressing FLAG-tagged TrkB were biotinylated for 30 min at 4°C. Endocytosis of biotinylated TrKB receptor was then induced by incubation with BDNF ligand. Internalized TrkB receptor was precipitated with streptavidin beads and quantified by immunoblotting with an anti-FLAG antibody.

2017-10-13 · The monoclonal and polyclonal anti-STxB antibodies were obtained as described previously (Johannes et al., 1997; Mallard et al., 1998). The Fab-fragment of 13C4 was prepared according to the manufacturer's instructions with the use of a commercial kit (Pierce, Rockford, IL).

Sorting nexin 8 regulates endosome-to-Golgi transport. Biochemical and Biophysical Research Communications, 2009. Audrun Utskarpen