Polyclonal rabbit anti-human fibrinogen antibody was from DAKOCytomation (Carpinteria, CA). Monoclonal anti-fibrin(ogen) antibody (59D8) was a generous gift of Drs. Marschall Runge (University of North Carolina [UNC]), Charles Esmon (Oklahoma College of Medicine), and Rodney Camire (University of Pennsylvania). Mouse anti-human γ’ chain-specific antibody (2.G2.H9) was from Millipore (Temecula, CA). Biotinylated secondary antibodies were from Vector Laboratories (Burlingame, CA). The AlexaFluor-488 protein labeling kit and 10% pre-cast Tris-glycine gels were from Invitrogen (Carlsbad, CA). Human α-thrombin and murine thrombin were from Enzyme Research Laboratories (South Bend, IN). Lipidated tissue factor (TF, Innovin) was from Siemens (Newark, DE). Phospholipid vesicles (phosphatidylserine/phosphatidylcholine/phosphatidylethanolamine) were prepared as described [26 (link)]. Bovine serum albumin was from Sigma-Aldrich (St. Louis, MO). Peroxidase substrate was from KPL (Gaithersburg, MD).
Polyclonal rabbit anti human fibrinogen antibody
Manufactured by Agilent Technologies
Sourced in United States
The Polyclonal rabbit anti-human fibrinogen antibody is a laboratory reagent that detects the presence of human fibrinogen, a blood plasma protein involved in blood clotting. This antibody is produced in rabbits and recognizes multiple epitopes on the fibrinogen molecule.
Automatically generated - may contain errors
Lab products found in correlation
Murine thrombin, by Enzyme Research (3 mentions)
Human α thrombin, by Enzyme Research (3 mentions)
Biotinylated secondary antibody, by Vector Laboratories (1 mentions)
Alexa fluor 488 protein labeling kit, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Ecl western blotting detection reagent, by GE Healthcare (1 mentions)
Chemidoc xrs plus, by Bio-Rad (1 mentions)
Tryptic soy broth tsb, by BD (1 mentions)
Tryptic soy agar, by BD (1 mentions)
Fxiiia, by Prolytix (1 mentions)
3 protocols using polyclonal rabbit anti human fibrinogen antibody
1
Quantification of Fibrinogen and Thrombin Interactions
2
Analyzing Plasma Fibrinogen Subunits via Western Blot
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The patient’s plasma fibrinogen and Aα, Bβ, and γ chains were determined using Western blotting. Purified normal plasma fibrinogen and normal plasma were used as normal controls. The patient’s and normal plasma were diluted 1000 times in N-[2-hydroxyethyl] piperazine-N’-[2-ethanesulfonic acid] (HEPES), pH 7.4, and 0.12 M NaCl buffer (HBS buffer), separated by SDS-PAGE in non-reducing conditions in a 10% polyacrylamide gel or reducing conditions in a 7% polyacrylamide gel, transferred into a nitrocellulose membrane, and developed with a polyclonal rabbit anti-human fibrinogen antibody (Dako, Carpinteria, CA, USA). The reacting species were visualized using a horseradish peroxidase (HRP) conjugated-goat anti-rabbit IgG antibody (Medical and Biological Laboratories, Nagoya, Japan). Blots were exposed using ECL Western Blotting Detection Reagents (GE Healthcare, Tokyo, Japan) and detected with a ChemiDoc XRS Plus (Bio-Rad, Hercules, CA, USA).
3
GBS Fibronectin-Binding Protein Assays
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Chemicals used in this study were purchased from Sigma-Aldrich, unless stated otherwise. Commercial antibodies used in this study included the anti-fibronectin antibody (clone FBN11, Invitrogen), anti-FXIIIA antibody (clone ab1834, Abcam), anti-transferrin antibody (clone PA3-913, Thermo Scientific), polyclonal rabbit anti-human fibrinogen antibody (code A0080, Dako), and anti-streptococcus group B antibody (catalog ab53584, Abcam). Tridegin was synthesized in house following procedures previously described (35 (link), 70 (link)), and FXIIIA was purchased from Haematologic Technologies.
The WT GBS strains used in this study were clinical isolates obtained from human newborns; these included A909, a capsular serotype Ia strain (71 (link)), and COH1, a capsular serotype III hypervirulent ST-17 clone (72 (link)). GBS were grown using tryptic soy broth (TSB) or tryptic soy agar (TSA) (Difco Laboratories) at 30°C or 37°C in 5% CO2. GBS mutants deficient for fibronectin-binding protein ScpB (ΔscpB, ref. 37 (link)), SfbA (ΔsfbA, ref. 38 (link)), and Fib1 (Δfib1, ref. 39 (link)) were previously described. The complemented pScpB plasmid was previously constructed (37 (link)).
The WT GBS strains used in this study were clinical isolates obtained from human newborns; these included A909, a capsular serotype Ia strain (71 (link)), and COH1, a capsular serotype III hypervirulent ST-17 clone (72 (link)). GBS were grown using tryptic soy broth (TSB) or tryptic soy agar (TSA) (Difco Laboratories) at 30°C or 37°C in 5% CO2. GBS mutants deficient for fibronectin-binding protein ScpB (ΔscpB, ref. 37 (link)), SfbA (ΔsfbA, ref. 38 (link)), and Fib1 (Δfib1, ref. 39 (link)) were previously described. The complemented pScpB plasmid was previously constructed (37 (link)).
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