Mice were intraperitoneally injected with anti-CD41 antibody (BD Bioscience) either daily at 0.1 mg/kg body weight in PBS or on day 0 and 3 only each at 0.5 mg/kg body weight. Control mice were intraperitoneally injected with 400 μL PBS in parallel. Platelet counts were analyzed on a HemaTrue veterinary hematology analyzer (Heska Corporation) with 50 μL whole blood collected from the retro-orbital venous plexus into EDTA-coated microtainer tubes (BD Bioscience)
Anti cd41 antibody
Manufactured by BD
Sourced in United States
The Anti-CD41 antibody is a laboratory reagent used for the detection and analysis of the CD41 protein, which is a component of the platelet glycoprotein IIb/IIIa complex. The antibody can be utilized in various immunoassay techniques, such as flow cytometry, to study the expression and distribution of CD41 in different cell types.
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6 protocols using anti cd41 antibody
1
Anti-CD41 Antibody-Induced Thrombocytopenia
2
Modelling Dengue Hemorrhagic Fever in Mice
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Male C57BL/6 mice were purchased from National Cheng Kung University (NCKU) Laboratory Animal Center and used at 6–8 weeks of age. ProT Tg mice were generated on a C57BL/6 background in our laboratory.72 (link) All animal work was conducted in animal biosafety level 2 facilities at NCKU. The experimental protocols adhered to the rules of the Animal Protection Act of Taiwan and were approved by the Animal Care and Use Committee of NCKU (IACUC number: 106120).
A two-hit model for dengue-induced hemorrhage was established as previously described.29 (link) Human primary infection with DENV can induce mild symptoms, known as dengue fever. However, secondary infection may induce life-threatening dengue hemorrhagic fever. The two-hit model in mice using DENV and anti-CD41 (PLT marker) antibody resembles human dengue infection. The first hit mimics viremia by injecting DENV, and the second hit imitates NS1-elicited autoantibody by offering anti-CD41 neutralizing antibody. ProT Tg and WT C57BL/6 mice were injected subcutaneously with 107 PFU of DENV-2 (PL046 strain) with or without injection of 1 mg/kg of anti-CD41 antibody (BD Biosciences, San Diego, CA, USA) after 24 h at the same site of DENV injection. Various parameters of mice were determined at different time points after viral infection.
A two-hit model for dengue-induced hemorrhage was established as previously described.29 (link) Human primary infection with DENV can induce mild symptoms, known as dengue fever. However, secondary infection may induce life-threatening dengue hemorrhagic fever. The two-hit model in mice using DENV and anti-CD41 (PLT marker) antibody resembles human dengue infection. The first hit mimics viremia by injecting DENV, and the second hit imitates NS1-elicited autoantibody by offering anti-CD41 neutralizing antibody. ProT Tg and WT C57BL/6 mice were injected subcutaneously with 107 PFU of DENV-2 (PL046 strain) with or without injection of 1 mg/kg of anti-CD41 antibody (BD Biosciences, San Diego, CA, USA) after 24 h at the same site of DENV injection. Various parameters of mice were determined at different time points after viral infection.
3
Flow Cytometry Analysis of Megakaryocyte Differentiation
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K562 cells were induced to differentiate with TPA (10 nM ) for hours as indicated. The treated cells were washed with phosphate-buffered saline and stained with phycoerythrin-conjugated anti-CD61 antibody or anti-CD41 antibody (BD Biosciences, San Jose, CA, USA). Differentiation of primary mouse megakaryocytes was performed by staining cells with fluorescently labeled anti-CD41 (cat. no. 17-0411, eBioscience, San Diego, CA, USA) or anti-CD42 (cat. no. 12-0421-82, eBioscience) antibodies or by staining DNA with 4,6-diamidinio-2-phenylindole (DAPI, 1 mg/ml). FACS data were acquired with a FACSAria instrument (BD Biosciences, Mountain View, CA, USA) and analyzed with FlowJo software (TreeStar, Ashland, OR, USA). All flow cytometry data depict the representative results from three independent experiments with duplicates.
4
Murine Models of Thrombocytopenia
5
Thrombocytopenia Induction for Photothrombosis
6
Platelet Aggregation Assay after Tail Injury
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At 20 min before the end of the experiment, the distal end of the tail (at 3 mm diameter) was cut off using a double guillotine cigar cutter. 22 The tail was submerged in pre-warmed (37 C) NaCl 0.9% (50 ml), and bleeding time was recorded. Platelet aggregation was measured using flow cytometry, as described. 23 In short, whole blood was incubated for 15 min with either anti-CD41 antibody or PE anti-CD61 antibody (BD Biosciences). Excess antibody was removed by two washing steps. The two stained and washed conditions were mixed and incubated at 37 C for 15 min with ADP 20 mM or protease-activated receptor 4 activating peptide 100 mM (Cayman Chemical, Ann Arbor, MI, USA) while aggregating at 600 rpm. In both activation and aggregation assays, we focused on ADP activation and ADP/PAR4-induced aggregation based on the platelet dysfunction seen after traumatic injury. 5 All flow cytometry analyses were performed using the FACS-Canto™ system (BD Biosciences). Antibody panels can be found in Supplementary Table S1.
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