FlexAble 2.0 CoraLite® Plus 647 Antibody Labeling Kit for Rabbit IgG

Application

IF, FC, WB

Reactivity

Rabbit

Conjugate

CoraLite® Plus 647

Cat no : KFA503

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Validation Data Gallery

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  Immunofluorescence of Mouse colon: FFPE Mouse colon sections were stained with anti-FUS (11570-1-AP) labeled with FlexAble 2.0 CoraLite® Plus 647 Kit (KFA503, magenta), anti-α-SMA (67735-1-Ig) labeled with FlexAble 2.0 CoraLite® Plus 555 Kit (KFA522, orange), anti-E-cadherin (60335-1-Ig) labeled with FlexAble 2.0 CoraLite® Plus 488 Kit (KFA561, green) and DAPI (blue).

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  Immunofluorescence of Mouse colon: FFPE Mouse colon sections were stained with anti-FUS (11570-1-AP) labeled with FlexAble 2.0 CoraLite® Plus 647 Kit (KFA503, magenta), anti-α-SMA (67735-1-Ig) labeled with FlexAble 2.0 CoraLite® Plus 555 Kit (KFA522, orange), anti-E-cadherin (60335-1-Ig) labeled with FlexAble 2.0 CoraLite® Plus 488 Kit (KFA561, green) and DAPI (blue).

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  Immunofluorescence of Mouse colon: FFPE Mouse colon sections were stained with anti-FUS (11570-1-AP) labeled with FlexAble 2.0 CoraLite® Plus 647 Kit (KFA503, magenta), anti-α-SMA (67735-1-Ig) labeled with FlexAble 2.0 CoraLite® Plus 555 Kit (KFA522, orange) and DAPI (blue).

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  Immunofluorescence of Mouse colon: FFPE Mouse colon sections were stained with anti-FUS (11570-1-AP) labeled with FlexAble 2.0 CoraLite® Plus 647 Kit (KFA503, magenta), anti-α-SMA (67735-1-Ig) labeled with FlexAble 2.0 CoraLite® Plus 555 Kit (KFA522, orange) and DAPI (blue).

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  Immunofluorescence of Rat brain: FFPE Rat brain sections were stained with anti-MAP2 (17490-1-AP) labeled with FlexAble 2.0 CoraLite® Plus 647 Kit (KFA503, magenta), anti-GFAP (60190-1-Ig) labeled with FlexAble 2.0 CoraLite® Plus 488 Kit (KFA541, green), anti-TDP-43 (67345-1-Ig) labeled with FlexAble 2.0 CoraLite® Plus 555 Kit (KFA542,organge) and DAPI (blue).

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  Immunofluorescence of Rat brain: FFPE Rat brain sections were stained with anti-MAP2 (17490-1-AP) labeled with FlexAble 2.0 CoraLite® Plus 647 Kit (KFA503, magenta), anti-GFAP (60190-1-Ig) labeled with FlexAble 2.0 CoraLite® Plus 488 Kit (KFA541, green), anti-TDP-43 (67345-1-Ig) labeled with FlexAble 2.0 CoraLite® Plus 555 Kit (KFA542,organge) and DAPI (blue).

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  Immunofluorescence of Hela cells: PFA-fixed cells were stained with rabbit IgG anti-Lamin antibody (12987-1-AP) labeled with FlexAble 2.0 CoraLite® Plus 555 Kit (KFA502, yellow), rabbit IgG anti-GM130 antibody (11308-1-AP) labeled with FlexAble 2.0 CoraLite® Plus 647 Kit (KFA503, magenta) and rabbit IgG anti-Tubulin antibody (80713-1-RR) labeled with FlexAble 2.0 CoraLite® Plus 750 Kit (KFA504, grey). Cell nuclei were stained with DAPI (blue).  Confocal images were acquired with a 63x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

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  Immunofluorescence of Hela cells: PFA-fixed cells were stained with rabbit IgG anti-Lamin antibody (12987-1-AP) labeled with FlexAble 2.0 CoraLite® Plus 555 Kit (KFA502, yellow), rabbit IgG anti-GM130 antibody (11308-1-AP) labeled with FlexAble 2.0 CoraLite® Plus 647 Kit (KFA503, magenta) and rabbit IgG anti-Tubulin antibody (80713-1-RR) labeled with FlexAble 2.0 CoraLite® Plus 750 Kit (KFA504, grey). Cell nuclei were stained with DAPI (blue).  Confocal images were acquired with a 63x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

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  Immunofluorescence of Hela cells: PFA-fixed cells were stained with rabbit IgG anti-TDP43 antibody directly conjugated with CoraLite® Plus 488 (CL488-10782, green), rabbit IgG anti-GM130 antibody directly conjugated with CoraLite® Plus 555 (CL555-11308, red), rabbit IgG anti-Lamin antibody (12987-1-AP) labeled with FlexAble 2.0 CoraLite® Plus 647 Kit (KFA503, magenta) and rabbit IgG anti-Tubulin antibody (80713-1-RR) labeled with FlexAble 2.0 CoraLite® Plus 750 Kit (KFA504, cyan). Cell nuclei were stained with DAPI (blue).  Confocal images were acquired with a 63x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

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  Immunofluorescence of Hela cells: PFA-fixed cells were stained with rabbit IgG anti-TDP43 antibody directly conjugated with CoraLite® Plus 488 (CL488-10782, green), rabbit IgG anti-GM130 antibody directly conjugated with CoraLite® Plus 555 (CL555-11308, red), rabbit IgG anti-Lamin antibody (12987-1-AP) labeled with FlexAble 2.0 CoraLite® Plus 647 Kit (KFA503, magenta) and rabbit IgG anti-Tubulin antibody (80713-1-RR) labeled with FlexAble 2.0 CoraLite® Plus 750 Kit (KFA504, cyan). Cell nuclei were stained with DAPI (blue).  Confocal images were acquired with a 63x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

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  Immunofluorescence of Hela cells: PFA-fixed cells were stained with rabbit IgG anti-GM130 antibody (11308-1-AP) labeled with FlexAble 2.0 CoraLite® Plus 647 Kit (KFA503, magenta). Cell nuclei were stained with DAPI (blue).  Confocal images were acquired with a 63x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

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  1X10^6 HeLa cells were intracellularly stained with anti-Mitofilin rabbit polyclonal antibody (10179-1-AP, red) or with isotype control antibody (30000-0-AP, blue) labeled with FlexAble 2.0 CoraLite® Plus 647 Kit (KFA503). Cells were fixed and permeabilized with Foxp3 /Transcription Factor Staining Buffer Kit (PF00011).

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  1X10^6 HeLa cells were intracellularly stained with anti-TOM20 rabbit polyclonal antibody (11802-1-AP, orange) or with isotype control antibody (30000-0-AP, blue) labeled with FlexAble 2.0 CoraLite® Plus 647 Kit (KFA503). Cells were fixed and permeabilized with Foxp3 /Transcription Factor Staining Buffer Kit (PF00011).

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Product Information

Product name	FlexAble 2.0 CoraLite® Plus 647 Antibody Labeling Kit for Rabbit IgG
Assay type	Antibody labeling
Tested applications	IF, FC, WB
Species Reactivity	Rabbit IgG
Antibody amount per labeling reaction	0.5 µg antibody
Conjugate	CoraLite® Plus 647
Excitation / Emission maxima wavelengths	654 nm/ 674 nm

Component	10 rxns	50 rxns	200 rxns
CoraLite® Plus 647 FlexLinker 2.0 for Rabbit IgG	10 μL	50 μL	200 μL
FlexQuencher for Rabbit IgG	20 μL	100 μL	400 μL
FlexBuffer	100 μL	500 μL	2000 μL

Storage

Store for 1 year at -20°C or for 6 months at +4°C after shipment.  Avoid exposure to light.

Others

Q: What is the difference between FlexAble and FlexAble 2.0 products?

A: FlexAble 2.0 has been improved to allow for more dyes per FlexLinker than the previous version of the kit.  This results in an increased signal per antibody.

Q: What are the FlexLinker, FlexQuencher and FlexBuffer?

A: The FlexLinker is a small polypeptide to which dyes are covalently conjugated that can label unconjugated primary antibodies. The FlexQuencher is an Fc-containing fragment that neutralizes the excess FlexLinker. The FlexBuffer is a PBS-based buffer.

Q: What is the largest quantity I can label?

A: With a standard kit size (50 reactions), you can label 25 µg of one antibody or up to 50 different antibodies. You can easily scale up the antibody amount per labeling approach.

Q: What is the lowest concentration of my primary antibody that I can use?

A: Our protocol uses 0.5 µg of primary antibody in 7 µL, which ends up at 0.07 mg/mL. If the concentration of your antibody is lower, you can also use a larger volume than 7 µL.

Q: Can I label primary antibodies stored in BSA, glycerol, Tris buffer and/or preservatives?

A: Yes, FlexAble Antibody Labeling Kits have been validated with carriers and amine buffers. Neither BSA nor amine buffers, in any chosen concentration, interfere with the labeling. 50% glycerol as well as preservatives like sodium azide are also compatible with the kit.

Q: How many different primary antibodies can I label with one kit?

A: You can label up to 50 different antibodies with our FlexAble 50 rxn Kit, and up to 10 antibodies with our FlexAble 10 rxn Kit.

Q: Will I observe cross-reactivity/leaking when I use two FlexAble-labeled antibodies from the same species during multiplexing?

A: FlexAble labels primary antibodies with a high affinity FlexLinker. Dissociation of FlexLinker from one antibody and association to another antibody is rare. If you observe leaking, we recommend adding more FlexQuencher to remove unbound FlexLinker, or you can try sequential staining of the labeled antibodies. 

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