Alexa fluor647 labelled goat anti human igg secondary ab

Manufactured by Jackson ImmunoResearch

Alexa Fluor647-labelled Goat Anti-human IgG secondary Ab is a fluorescently-labelled secondary antibody. It is designed to detect and bind to human immunoglobulin G (IgG) antibodies. The Alexa Fluor647 dye provides a bright red fluorescent signal that can be detected and measured using appropriate instrumentation.

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Lab products found in correlation

96 well round bottom plate, by Corning (3 mentions) Expifectamine cho reagent, by Thermo Fisher Scientific (3 mentions) Ze5 cytometer, by Bio-Rad (3 mentions) Optipro sfm, by Thermo Fisher Scientific (1 mentions)

3 protocols using alexa fluor647 labelled goat anti human igg secondary ab

Investigating SARS-CoV-2 Spike Glycoprotein Variants

Cited 1 time

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ExpiCHO-S cells were seeded at 6 × 10⁶ cells/mL in a volume of 5 mL in a 50 mL bioreactor. The following day, cells were transfected with SARS-CoV-2 spike glycoprotein-encoding pcDNA3.1(+) plasmids (BetaCoV/Wuhan-Hu-1/2019, accession number MN908947, Wuhan D614; Omicron BA.1 and BA.2 generated by overlap PCR mutagenesis of the Wuhan D614 plasmid) harboring the Δ19 C-terminal truncation^{27 (link)}. Spike encoding plasmids were diluted in cold OptiPRO SFM (Life Technologies, 12309-050), mixed with ExpiFectamine CHO Reagent (Life Technologies, A29130) and added to cells. Transfected cells were then incubated at 37˚C with 8% CO₂ with an orbital shaking speed of 250 RPM (orbital diameter of 25 mm) for 24 to 48 h. Transiently transfected ExpiCHO-S cells were harvested and washed twice in wash buffer (PBS 2% FBS, 2 mM EDTA). Cells were counted and distributed into round bottom 96-well plates (Corning, 3799) and incubated with serial dilutions of mAb starting at 10 μg/mL. Alexa Fluor647-labelled Goat Anti-human IgG secondary Ab (Jackson ImmunoResearch, 109–606–098) was prepared at 2 μg/mL and added onto cells after two washing steps. Cells were then washed twice and resuspended in wash buffer for data acquisition at ZE5 cytometer (BioRad).

Case J.B., Mackin S., Errico J.M., Chong Z., Madden E.A., Whitener B., Guarino B., Schmid M.A., Rosenthal K., Ren K., Dang H.V., Snell G., Jung A., Droit L., Handley S.A., Halfmann P.J., Kawaoka Y., Crowe JE J.r., Fremont D.H., Virgin H.W., Loo Y.M., Esser M.T., Purcell L.A., Corti D, & Diamond M.S. (2022). Resilience of S309 and AZD7442 monoclonal antibody treatments against infection by SARS-CoV-2 Omicron lineage strains. Nature Communications, 13, 3824.

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Quantifying SARS-CoV-2 Spike Protein Binding

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ExpiCHO-S cells were seeded at 6 × 10⁶ cells cells/mL
in a volume of 5 mL in a 50 mL bioreactor. Spike coding plasmids were diluted in
cold OptiPRO SFM, mixed with ExpiFectamine CHO Reagent (Life Technologies) and
added to the cells. Transfected cells were then incubated at 37°C with 8%
CO₂ with an orbital shaking speed of 120 RPM (orbital diameter of
25 mm) for 42 hours. Transiently transfected ExpiCHO-S cells were harvested and
washed two times in wash buffer (PBS 1% BSA, 2 mM EDTA). Cells were counted and
distributed into round bottom 96-well plates (Corning) and incubated with 10
μg/mL S2H97, S2X35 or S309 mAb. Alexa Fluor647-labelled Goat Anti-Human
IgG secondary Ab (Jackson ImmunoResearch 109-607-003) was prepared at 1.5
μg/mL added onto cells after two washing steps. Cells were then washed
twice and resuspended in wash buffer for data acquisition on a ZE5 cytometer
(Biorad).

Starr T.N., Czudnochowski N., Liu Z., Zatta F., Park Y.J., Addetia A., Pinto D., Beltramello M., Hernandez P., Greaney A.J., Marzi R., Glass W.G., Zhang I., Dingens A.S., Bowen J.E., Tortorici M.A., Walls A.C., Wojcechowskyj J.A., De Marco A., Rosen L.E., Zhou J., Montiel-Ruiz M., Kaiser H., Dillen J., Tucker H., Bassi J., Silacci-Fregni C., Housley M.P., di Iulio J., Lombardo G., Agostini M., Sprugasci N., Culap K., Jaconi S., Meury M., Dellota E., Abdelnabi R., Foo S.Y., Cameroni E., Stumpf S., Croll T.I., Nix J.C., Havenar-Daughton C., Piccoli L., Benigni F., Neyts J., Telenti A., Lempp F.A., Pizzuto M.S., Chodera J.D., Hebner C.M., Virgin H.W., Whelan S.P., Veesler D., Corti D., Bloom J.D, & Snell G. (2021). SARS-CoV-2 RBD antibodies that maximize breadth and resistance to escape. Nature, 597(7874), 97-102.

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ExpiCHO-S Cell SARS-CoV-2 Spike Binding

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ExpiCHO-S cells were seeded at 6 × 10⁶ cells cells/mL in a volume of 5 mL in a 50 mL bioreactor. Spike coding plasmids were diluted in cold OptiPRO SFM, mixed with ExpiFectamine CHO Reagent (Life Technologies) and added to the cells. Transfected cells were then incubated at 37°C with 8% CO₂ with an orbital shaking speed of 120 RPM (orbital diameter of 25 mm) for 42 hours. Transiently transfected ExpiCHO cells were harvested and washed two times in wash buffer (PBS 1% BSA, 2 mM EDTA). Cells were counted and distributed into round bottom 96-well plates (Corning) and incubated with 10 μg/mL S2H97, S2X35 or S309 mAb. Alexa Fluor647-labelled Goat Anti-Human IgG secondary Ab (Jackson ImmunoResearch 109-607-003) was prepared at 1.5 mg/mL added onto cells after two washing steps. Cells were then washed twice and resuspended in wash buffer for data acquisition at ZE5 cytometer (Biorad).

Starr T.N., Czudnochowski N., Zatta F., Park Y.J., Liu Z., Addetia A., Pinto D., Beltramello M., Hernandez P., Greaney A.J., Marzi R., Glass W.G., Zhang I., Dingens A.S., Bowen J.E., Wojcechowskyj J.A., De Marco A., Rosen L.E., Zhou J., Montiel-Ruiz M., Kaiser H., Tucker H., Housley M.P., di Iulio J., Lombardo G., Agostini M., Sprugasci N., Culap K., Jaconi S., Meury M., Dellota E., Cameroni E., Croll T.I., Nix J.C., Havenar-Daughton C., Telenti A., Lempp F.A., Pizzuto M.S., Chodera J.D., Hebner C.M., Whelan S.P., Virgin H.W., Veesler D., Corti D., Bloom J.D, & Snell G. (2021). Antibodies to the SARS-CoV-2 receptor-binding domain that maximize breadth and resistance to viral escape. bioRxiv.

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