MDALDL or OVA (10 μg/ml) was incubated at 4°C for 18–24 h with affinity purified rabbit anti-MDA IgG or rabbit anti-OVA IgG at 30 μg/ml to prepare soluble MDALDL-IC and OVA-IC. Soluble IC was centrifuged at 10000 rpm for 20 min to remove any particulate IC. To prepare insoluble BSA-IC, IgG-free BSA (10 μg/ml in PBS, Sigma A0281) was incubated with rabbit anti-BSA antiserum (300 μl of 2 mg/ml anti-BSA IgG, Sigma B1520) for 20 min at 37°C. Insoluble BSA-IC was centrifuged at 5000 rpm for 2 min and pellet containing insoluble BSA-IC was washed with PBS three times and resuspended in 310 μl serum free RPMI medium. BSA-IC (15 μl/1 ×106 cells/well) was used to activate macrophages. In all the assays both soluble and insoluble ICs were prepared fresh and used within 48 h.
Anti bsa igg
Manufactured by Merck Group
Sourced in Spain
Anti-BSA IgG is a laboratory reagent used for the detection and quantification of bovine serum albumin (BSA) in various samples. It is a highly specific antibody that binds to BSA, allowing for its identification and measurement.
Automatically generated - may contain errors
Lab products found in correlation
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
Polyethersulfone membrane, by Merck Group (1 mentions)
Human serum albumin (hsa), by Merck Group (1 mentions)
Alexa 488 conjugated α rabbit igg, by Thermo Fisher Scientific (1 mentions)
Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions)
Interferon γ, by Thermo Fisher Scientific (1 mentions)
Il 13, by Thermo Fisher Scientific (1 mentions)
3 protocols using anti bsa igg
1
Preparation and Use of Immune Complexes
2
Immobilization of Proteins on Glass Slides
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Sodium phosphate buffer
(PBS, 8 mM Na2HPO4, 2 mM, 137 mM NaCl, 2.7 mM
KCl, pH 7.4), PBS-T (PBS with polysorbate 20 0.05% v/v), and carbonate–bicarbonate
buffer (15 mM Na2CO3, 34 mM NaHCO3, pH 9.6) were prepared with purified water (Milli-Q, Millipore Iberica,
Darmstadt, Germany) and filtered through 0.2 μm polyethersulfone
membranes (Merck, Darmstadt, Germany). BSA, polysorbate 20 (Tween
20), anti-BSA IgG produced in rabbit (whole antiserum), human serum
albumin (HSA), human IgG, hemoglobin, goat anti-rabbit antibodies
labeled with 5 nm gold nanoparticles, and silver enhancers were supplied
by Sigma-Aldrich (Madrid, Spain). An Alexa Fluor 647 conjugation kit
was from Abcam (Cambridge, United Kingdom). Polydimethylsiloxane (PDMS)
Sylgard 184 was supplied by Dow Corning (Wiesbaden, Germany). Human
serum obtained by centrifugation of a pool of blood samples (type
AB) from male donors was provided by Sigma-Aldrich (Madrid, Spain).
Glass slides (25 × 75 × 1 mm) were purchased from Labbox
(Barcelona, Spain).
Glass slides were washed three times by
sonication in ethanol (30% in water, 5 min) and dried under a stream
of air. Then, protein solutions in carbonate buffer (500 μL,
25 μg·mL–1) were incubated overnight
on the glass slides at 4 °C (Figure S1 ). Finally, glass slides were rinsed with deionized water and dried
by air stream.
(PBS, 8 mM Na2HPO4, 2 mM, 137 mM NaCl, 2.7 mM
KCl, pH 7.4), PBS-T (PBS with polysorbate 20 0.05% v/v), and carbonate–bicarbonate
buffer (15 mM Na2CO3, 34 mM NaHCO3, pH 9.6) were prepared with purified water (Milli-Q, Millipore Iberica,
Darmstadt, Germany) and filtered through 0.2 μm polyethersulfone
membranes (Merck, Darmstadt, Germany). BSA, polysorbate 20 (Tween
20), anti-BSA IgG produced in rabbit (whole antiserum), human serum
albumin (HSA), human IgG, hemoglobin, goat anti-rabbit antibodies
labeled with 5 nm gold nanoparticles, and silver enhancers were supplied
by Sigma-Aldrich (Madrid, Spain). An Alexa Fluor 647 conjugation kit
was from Abcam (Cambridge, United Kingdom). Polydimethylsiloxane (PDMS)
Sylgard 184 was supplied by Dow Corning (Wiesbaden, Germany). Human
serum obtained by centrifugation of a pool of blood samples (type
AB) from male donors was provided by Sigma-Aldrich (Madrid, Spain).
Glass slides (25 × 75 × 1 mm) were purchased from Labbox
(Barcelona, Spain).
Glass slides were washed three times by
sonication in ethanol (30% in water, 5 min) and dried under a stream
of air. Then, protein solutions in carbonate buffer (500 μL,
25 μg·mL–1) were incubated overnight
on the glass slides at 4 °C (
by air stream.
3
Immunofluorescence Labeling of BSA
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Highly purified BSA (Cat # A0281) was purchased from Sigma. Interferon-γ (Cat # 315–05; Lot # 061398), IL4 (Cat # 214–14; Lot # 111249), and IL13 (Cat # 210–13; Lot # 111207) were purchased from Peprotech. anti-BSA IgG (Cat #B1520) was purchased from Sigma. Alexa 488-conjugated α-rabbit IgG (Cat # A11070) was from Invitrogen Life Technologies.
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