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Alexa fluor 488 conjugated anti ratigg h l

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Alexa-Fluor-488-conjugated anti-ratIgG (H+L) is a secondary antibody that binds to the heavy and light chains of rat immunoglobulin G (IgG). The antibody is conjugated to Alexa Fluor 488, a fluorescent dye that emits green fluorescence when excited by a 488 nm light source. This product can be used in various immunochemical techniques that require the detection of rat IgG.

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Lab products found in correlation

Lsm 880 confocal microscope, by Zeiss (6 mentions) Anti gfp clone fm264g, by BioLegend (3 mentions) Vectashield antifade mounting medium, by Vector Laboratories (3 mentions) Concavity microscope slide, by Thermo Fisher Scientific (2 mentions) Anti cd4 clone rm4 5, by BioLegend (2 mentions) Anti cd8a clone 53 6.7, by BioLegend (2 mentions) Anti mdll4 clone hmd4 1, by BioLegend (2 mentions) Antibodiesalexafluor 594 conjugated anti rat igg h l, by Jackson ImmunoResearch (2 mentions) Anti hamster biotin, by Jackson ImmunoResearch (2 mentions) Alexafluor 594 conjugated streptavidin, by Jackson ImmunoResearch (2 mentions) Airyscan andzen software, by Zeiss (2 mentions) Anti cd3 clone 145 2c11, by BioLegend (2 mentions) Anti cd3 clone ucht1, by BioLegend (1 mentions) Alexafluor 594 conjugated anti mouse igg h l, by Jackson ImmunoResearch (1 mentions) Airyscan, by Zeiss (1 mentions) Fv10c o, by Olympus (1 mentions)

4 protocols using alexa fluor 488 conjugated anti ratigg h l

1

Immunofluorescence Staining of Antigen-Presenting Cells

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PSC-ATOs were fixed in 4% Formaldehyde (Sigma-Aldrich, St. Louis, MO) for 30 minutes at room temperature followed by
3× 10 min. washes in PBST (0.3% Triton X-100) and a 1 hour block in PBST/BSA (2% BSA). ATOs were stained with anti-GFP
(clone FM264G; Biolegend, San Diego, CA) at a 1:100 dilution and anti-CD3 (clone UCHT1; Biolegend, San Diego, CA) at a 1:50
dilution overnight at 4ºC. Secondary antibodies AlexaFluor-594-conjugated anti-mouse IgG (H+L) (Jackson ImmunoResearch,
West Grove, PA) and AlexaFluor-488-conjugated anti-rat IgG (H+L) (Jackson ImmunoResearch West Grove, PA) were added at a 1:200
dilution for 2 hours at room temperature. Each ATO was mounted individually in Vectashield Antifade Mounting Medium (Vector
Laboratories, England) on a concavity microscope slide (ThermoFisher Scientific, Grand Island, NY). Immunofluorescence images
were acquired on a Zeiss LSM 880 confocal microscope equipped with Airyscan and Zen software (Zeiss, Jena, Germany).
Montel-Hagen A., Seet C.S., Li S., Chick B., Zhu Y., Chang P., Tsai S., Sun V., Lopez S., Chen H.C., He C., Chin C.J., Casero D, & Crooks G.M. (2019). Organoid-induced differentiation of conventional T cells from human pluripotent stem cells. Cell stem cell, 24(3), 376-389.e8.
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2

Immunofluorescence Imaging of M-ATOs

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M-ATOs were fixed in 4% Formaldehyde (Sigma-Aldrich, Cat#
F8775) for 30 minutes at room temperature followed by 3×10
min washes in PBST (0.3% Triton X-100) and a 1-hour block in PBST/BSA (2%
BSA). M-ATOs were stained with anti-CD8a (clone 53–6.7;
Biolegend), anti-mDLL4 (clone HMD4–1;
Biolegend), anti-CD4 (clone RM4–5;
Biolegend), and anti-GFP (clone FM264G; Biolegend) at a
1:100 dilution, and anti-CD3 (clone 145–2C11; Biolegend)
at a 1:50 dilution overnight at 4°C. Secondary antibodies
AlexaFluor-594-conjugated anti-rat IgG (H+L) (Jackson ImmunoResearch,
Cat# 712–585-150) or Alexa-Fluor-488-conjugated anti-rat
IgG (H+L) (Jackson ImmunoResearch, Cat# 712–485-153)
were added at a 1:200 dilution for 2 hours at room temperature. For
anti-mDLL4, anti-hamster biotin (Jackson ImmunoResearch, Cat#
127–065-160) was added at a 1:500 dilution for 2 hours at
room temperature, and then AlexaFluor-594-conjugated Streptavidin
(Jackson ImmunoResearch, Cat# 016–580-084) was added
at a 1:800 dilution for 30 minutes at room temperature. Each M-ATO was
mounted individually in Vectashield Antifade Mounting Medium (Vector
Laboratories, Cat# H1000) on a concavity microscope slide
(Fisher Scientific). Immunofluo-rescence images were
acquired on a Zeiss LSM 880 confocal microscope equipped with Airyscan and
Zen software (Zeiss).
Montel-Hagen A., Sun V., Casero D., Tsai S., Zampieri A., Jackson N., Li S., Lopez S., Zhu Y., Chick B., He C., de Barros S.C., Seet C.S, & Crooks G.M. (2020). In Vitro Recapitulation of Murine Thymopoiesis from Single Hematopoietic Stem Cells. Cell reports, 33(4), 108320.
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3

Immunofluorescence Imaging of M-ATOs

Check if the same lab product or an alternative is used in the 5 most similar protocols
M-ATOs were fixed in 4% Formaldehyde (Sigma-Aldrich, Cat#
F8775) for 30 minutes at room temperature followed by 3×10
min washes in PBST (0.3% Triton X-100) and a 1-hour block in PBST/BSA (2%
BSA). M-ATOs were stained with anti-CD8a (clone 53–6.7;
Biolegend), anti-mDLL4 (clone HMD4–1;
Biolegend), anti-CD4 (clone RM4–5;
Biolegend), and anti-GFP (clone FM264G; Biolegend) at a
1:100 dilution, and anti-CD3 (clone 145–2C11; Biolegend)
at a 1:50 dilution overnight at 4°C. Secondary antibodies
AlexaFluor-594-conjugated anti-rat IgG (H+L) (Jackson ImmunoResearch,
Cat# 712–585-150) or Alexa-Fluor-488-conjugated anti-rat
IgG (H+L) (Jackson ImmunoResearch, Cat# 712–485-153)
were added at a 1:200 dilution for 2 hours at room temperature. For
anti-mDLL4, anti-hamster biotin (Jackson ImmunoResearch, Cat#
127–065-160) was added at a 1:500 dilution for 2 hours at
room temperature, and then AlexaFluor-594-conjugated Streptavidin
(Jackson ImmunoResearch, Cat# 016–580-084) was added
at a 1:800 dilution for 30 minutes at room temperature. Each M-ATO was
mounted individually in Vectashield Antifade Mounting Medium (Vector
Laboratories, Cat# H1000) on a concavity microscope slide
(Fisher Scientific). Immunofluo-rescence images were
acquired on a Zeiss LSM 880 confocal microscope equipped with Airyscan and
Zen software (Zeiss).
Montel-Hagen A., Sun V., Casero D., Tsai S., Zampieri A., Jackson N., Li S., Lopez S., Zhu Y., Chick B., He C., de Barros S.C., Seet C.S, & Crooks G.M. (2020). In Vitro Recapitulation of Murine Thymopoiesis from Single Hematopoietic Stem Cells. Cell reports, 33(4), 108320.
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4

Immunofluorescent Detection of LYVE-1 and CD31

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Mouse tissue sections prepared as described in the previous section were treated with combinations of anti‐LYVE‐1 (38M, 5 μg/mL) rat mAb and anti‐LYVE‐1 or anti‐CD31 rabbit pAb (1:100 diluted) overnight at RT. After extensive washing with PBS, tissue sections were treated with 1:200 diluted species‐specific Alexa Fluor 488‐conjugated anti‐rat IgG (H+L) and Alexa Fluor 647‐conjugated anti‐rabbit IgG (H+L) donkey pAb (Jackson ImmunoResearch) for 60 minutes at RT. The localization of antibody‐defined components was observed on confocal laser fluorescence microscopy with FV10C‐O (Olympus, Tokyo, Japan).
Hara Y., Torii R., Ueda S., Kurimoto E., Ueda E., Okura H., Tatano Y., Yagi H., Ohno Y., Tanaka T., Masuko K, & Masuko T. (2018). Inhibition of tumor formation and metastasis by a monoclonal antibody against lymphatic vessel endothelial hyaluronan receptor 1. Cancer Science, 109(10), 3171-3182.
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