Mycobacterial species M. malmoense, M. simiae, M. chelonae and M. smegmatis were grown in 7H9 culture media at 37 °C. Bacteria was harvested at OD600 of 0.8, washed and resuspended in 1X PBS to OD600 of 0.4. It was incubated with antibody HuMab-28-009 at a concentration of 5 µg/mL for 1 h at 4 °C. The bacteria were then washed with FACS buffer (1X PBS, 2% FBS, 0.1% sodium azide) and incubated with Alexa Fluor 647 anti-human IgG (Thermo Scientific Cat. No. A-21445) at 1:200 in FACS buffer for 30 min, followed by washing with FACS buffer to remove the unbound. The bacterial samples were then fixed with 4% formaldehyde solution for 15 min at room temperature and analysed on BD LSRFortessa flow cytometer (Data availability—Figshare 10.6084/m9.figshare.23550108).
Alexa fluor 647 anti human igg
Manufactured by Thermo Fisher Scientific
Alexa Fluor 647 anti-human IgG is a fluorescent-labeled secondary antibody that binds to human immunoglobulin G (IgG) antibodies. It is used to detect and quantify the presence of human IgG in various laboratory and research applications.
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Lab products found in correlation
3 protocols using alexa fluor 647 anti human igg
1
Mycobacterial Antibody Binding Assay
2
Mycobacterial Antibody Binding Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Mycobacterial species M. malmoense, M. simiae, M. chelonae and M. smegmatis were grown in 7H9 culture media at 37 °C. Bacteria was harvested at OD600 of 0.8, washed and resuspended in 1X PBS to OD600 of 0.4. It was incubated with antibody HuMab-28-009 at a concentration of 5 µg/mL for 1 h at 4 °C. The bacteria were then washed with FACS buffer (1X PBS, 2% FBS, 0.1% sodium azide) and incubated with Alexa Fluor 647 anti-human IgG (Thermo Scientific Cat. No. A-21445) at 1:200 in FACS buffer for 30 min, followed by washing with FACS buffer to remove the unbound. The bacterial samples were then fixed with 4% formaldehyde solution for 15 min at room temperature and analysed on BD LSRFortessa flow cytometer (Data availability—Figshare 10.6084/m9.figshare.23550108).
3
Mycobacterial Antibody Binding Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mycobacterial species M. malmoense, M. simiae, M. chelonae and M. smegmatis were grown in 7H9 culture media at 37 °C. Bacteria was harvested at OD600 of 0.8, washed and resuspended in 1X PBS to OD600 of 0.4. It was incubated with antibody HuMab-28-009 at a concentration of 5 µg/mL for 1 h at 4 °C. The bacteria were then washed with FACS buffer (1X PBS, 2% FBS, 0.1% sodium azide) and incubated with Alexa Fluor 647 anti-human IgG (Thermo Scientific Cat. No. A-21445) at 1:200 in FACS buffer for 30 min, followed by washing with FACS buffer to remove the unbound. The bacterial samples were then fixed with 4% formaldehyde solution for 15 min at room temperature and analysed on BD LSRFortessa flow cytometer (Data availability—Figshare 10.6084/m9.figshare.23550108).
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