ADAMTS13 activity was measured using the fluorescence resonance energy transfer substrate VWF73 (FRETS-VWF73) as previously described [18 (link)]. VWF antigen (VWF:Ag) levels were determined with an in-house enzyme-linked immunosorbent assay, using polyclonal rabbit anti-human VWF antibodies (DakoCytomation, Glostrup, Denmark) for catching and tagging. Fibrinogen levels were derived from the clotting curve of the prothrombin time assay using Thromborel S as a reagent on an automated coagulation laboratory (ACL 300, Instrumentation Laboratory). In a subset of 1208 participants of RS-I-3, an extended panel of inflammatory and immunology markers was measured as previously described, including complement, immunoglobulins, and cytokines [19 (link)].
Polyclonal rabbit anti human vwf antibody
Manufactured by Agilent Technologies
Sourced in Denmark
Polyclonal rabbit anti-human VWF antibodies are laboratory reagents used for the detection and analysis of von Willebrand factor (VWF) in biological samples. These antibodies are produced by immunizing rabbits with human VWF and then purifying the resulting polyclonal antibodies.
Automatically generated - may contain errors
Lab products found in correlation
Acl 300, by Werfen (1 mentions)
Lgt agarose type 7, by Merck Group (1 mentions)
Trypsin edta solution, by Thermo Fisher Scientific (1 mentions)
Streptavidin horseradish peroxidase, by Vector Laboratories (1 mentions)
3 3 diaminobenzidine dab chromogen, by Vector Laboratories (1 mentions)
Vectamount, by Vector Laboratories (1 mentions)
Maxisorp plate, by Thermo Fisher Scientific (1 mentions)
Ristocetin, by Merck Group (1 mentions)
Taq dna polymerase, by New England Biolabs (1 mentions)
Goat anti mouse igg, by Beckman Coulter (1 mentions)
Restriction enzyme, by New England Biolabs (1 mentions)
T4 dna ligase, by New England Biolabs (1 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
Triton x, by Merck Group (1 mentions)
Tyrode s buffer, by Merck Group (1 mentions)
P nitrophenyl phosphate, by Merck Group (1 mentions)
Ethylene glycol tetraacetic acid (egta), by Merck Group (1 mentions)
Mgcl2, by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
11 protocols using polyclonal rabbit anti human vwf antibody
1
ADAMTS13 Activity and VWF Levels Assessment
2
Measuring VWF and ADAMTS13 in Elderly Cohorts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Citrated plasma samples were collected at the third visit of RS-I and at the baseline examination of RS-II, and stored at −80°C. Between June and October 2013, we measured ADAMTS13 activity using a kinetic assay based on the fluorescence resonance energy transfer substrate VWF73 (FRETS-VWF73) assay [27 (link)]. Plasma samples were measured against a reference curve of serially diluted pooled normal human plasma defined as having an ADAMTS13 activity of 1 IU/ml, and we expressed ADAMTS13 activity as a percentage of this reference value. The ADAMTS13 activity of 6258 participants was measured: 3791 from RS-I and 2467 from RS-II.
Between July and October of 2008, VWF antigen levels (in IU/ml) were determined with an in-house ELISA using polyclonal rabbit anti-human VWF antibodies (DakoCytomation, Glostrop, Denmark) for capturing and tagging [28 (link)]. The intra-assay CV was 5.8% and the interassay CV was 7.8%. VWF antigen levels were measured in 3968 individuals from RS-I and in 2561 individuals from RS-II.
In total, 5176 participants with VWF and ADAMTS13 measurements also had a fasting glucose measurement and were free of diabetes at baseline, while 4232 participants were free of prediabetes at baseline (ESM Fig.2 and 3 ).
Between July and October of 2008, VWF antigen levels (in IU/ml) were determined with an in-house ELISA using polyclonal rabbit anti-human VWF antibodies (DakoCytomation, Glostrop, Denmark) for capturing and tagging [28 (link)]. The intra-assay CV was 5.8% and the interassay CV was 7.8%. VWF antigen levels were measured in 3968 individuals from RS-I and in 2561 individuals from RS-II.
In total, 5176 participants with VWF and ADAMTS13 measurements also had a fasting glucose measurement and were free of diabetes at baseline, while 4232 participants were free of prediabetes at baseline (ESM Fig.
3
VWF and ADAMTS13 Measurements in Plasma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fasting venous blood samples were taken at the research centre, and citrated plasma was stored at –80 °C. We determined VWF antigen with an in-house enzyme-linked immunosorbent assay using polyclonal rabbit antihuman VWF antibodies (DakoCytomation, Glostrop, Denmark) for catching and tagging. The intra-assay coefficient of variation was 5.8% and the inter-assay coefficient of variation was 7.8%. We measured ADAMTS13 activity using a kinetic assay based on the fluorescence resonance energy transfer substrate VWF73 (FRETSVWF73) assay19 (link). This assay uses a peptide containing the ADAMTS13 cleavage site of VWF, and thus captures variation in the VWF cleavage rate determined by ADAMTS13 levels and structure. Plasma samples were measured against a reference curve of serial dilutions of normal human plasma defined to have an ADAMTS13 activity of 1 IU/mL, and we expressed ADAMTS13 activity as a percentage of this. Ten percent of the samples were retested and all were within 25% variation. From these measurements, we also calculated the ratio between ADAMTS13 activity and VWF antigen levels.
4
Plasma Biomarkers Measurement Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Citrated blood was centrifuged at 2000g for 10 min; then the plasma was centrifuged at 14,000g for 10 min and stored in aliquots at -80 °C. VWF:Antigen (VWF:Ag) levels were measured with an in-house ELISA, using polyclonal rabbit anti-human VWF antibodies (Dakocytomation, Glostrup, Denmark) for catching and tagging. ADAMTS13 activity was measured using the Fluorescence Resonance Energy Transfer Substrate VWF 73 (FRETS-VWF73) [23] . Total fibrinogen levels were measured according to von Clauss on a fully-automated coagulation analyzer (Sysmex CS-5100 system, Siemens Healthcare Diagnostics, Breda, the Netherlands). Fibrinogen γ' antigen levels were measured with an enzyme-linked immunosorbent assay as described previously using anti-γ' fibrinogen antibodies for catching and HRP-labeled rabbit anti-fibrinogen antibodies for tagging [24] .
5
Immunohistochemical Analysis of Lung Vasculature
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
On P7 and P14, rat pups were anesthetized, and their lungs were harvested and prepared for immunohistochemistry and histological analyses, as previously described (14 (link),33 (link)). For immunohistochemical evaluation of von Willebrand factor (vWF), the deparaffinized lung sections were incubated with polyclonal rabbit antihuman vWF antibody (DakoCytomation, Glostrup, Denmark) and a biotin-labeled donkey antimouse secondary antibody (Jackson ImmunoResearch Laboratories, West Grove, PA). Immune complexes were visualized using diaminobenzidine (Vector Laboratories, Burlingame, CA). Light hematoxylin and eosin was applied as a counterstain.
6
SDS-agarose Electrophoresis for VWF Multimers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SDS-agarose discontinuous gel electrophoresis was performed as previously described [12] (link). Briefly, medium resolution (1.6%) gel (LGT agarose type VII, Sigma, Munich, Germany) was prepared and the samples were diluted according to their VWF content in sample buffer. Electrophoresis was performed for ∼18 h at 55 V. After electrophoresis, the multimers were transferred onto nitrocellulose membrane by electroblotting using transfer buffer (0.05 M phosphate, pH 7.4 with 0.04 M SDS, without methanol). After transfer, nonspecific binding sites were blocked with low-fat milk. All of the following incubation and washing steps were performed in low-fat milk. The membrane was incubated overnight at a 1∶2000 dilution of polyclonal rabbit anti-human VWF antibody (Dako, Glostrup, Denmark) or 1∶250 dilution of monoclonal mouse anti-human AAT type PiZ antibody (Clone ATZ11). Detection was performed using chemiluminescence (Fluorchem™, Alpha Innotech Corp., San Leandro, California).
7
Quantifying Angiogenesis in Tissue Scaffolds
8
Quantifying Plasma VWF Antigen by ELISA
9
Plasma biomarkers in Gastric Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The plasma control group consisted of 32 healthy subjects (15 females and 17 males) aged 21-63 years (average age: 42.2 ± 13.3). Plasma samples from the control group and the group of patients with GC were prepared by centrifuging anticoagulated blood (in 3.8 g/dL sodium citrate) specimens at 2,000 g for 15 min at 4°C, and stored in aliquots at -80°C until analysis. The plasma vWF activity was detected using a commercially available direct enzyme-linked immunosorbent assay (ELISA) kit (IMUBIND; American Diagnostica Inc., Stamford, CT, USA). The plasma vWF:Ag was quantified by sandwich ELISA using the rabbit anti-human vWF polyclonal antibody (Dako, Kyoto, Japan). Serum concentrations of VEGF were analyzed using a commercially available direct ELISA kit (NeoBioscience Technology Co. Ltd, Beijing, China).
10
Production and Characterization of Anti-VWF Monoclonal Antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood donations were from healthy volunteers who provided written informed consent. The pMH3 vector and CHO-S cells were purchased from AmProtein (Hangzhou, China). Restriction enzymes, T4 DNA ligase, and Taq DNA polymerase were obtained from New England BioLabs (Beverly, MA, USA). Rabbit anti-human VWF polyclonal antibody and rabbit anti-human VWF-HRP antibody were purchased from Dako Cytomation (Glostrup, Denmark). Ristocetin, tetramethylbenizidine (TMB), and collagen type III from human placenta were purchased from Sigma (St. Louis, MO, USA). Goat anti-mouse IgG and Goat anti-mouse IgG labeled with horseradish peroxidase (HRP) were purchased from Beckman-Coulter (Brea, CA, USA). SZ-123, a murine anti-human VWF A3 domain mAb (IgG1) was produced by standard hybridoma technology in our laboratory as described previously [22 (link)]. SZ-34, a murine mAb that binds to the VWF A2 domain but does not inhibit its function, was produced by standard hybridoma technology in our laboratory [25 (link)].
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!