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FlexAble CoraLite® Plus 488 Antibody Labeling Kit for Mouse IgG2a

Application

IF, FC, WB

Reactivity

Mouse

Conjugate

CoraLite® Plus 488

Cat No : KFA041

Print Protocol

Validation Data Gallery

  • Immunofluorescence of HeLa: PFA-fixed HeLa cells were stained with anti-PAF49 labeled with FlexAble CoraLite® Plus 488 Kit (KFA041, green), anti-Tubulin (66240-1-Ig) labeled with FlexAble CoraLite® Plus 555 Kit (KFA042, yellow), anti-Lamin labeled with FlexAble CoraLite® Plus 647 Kit (KFA043, magenta) and DAPI (blue). Confocal images were acquired with a 100x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

    Immunofluorescence of HeLa: PFA-fixed HeLa cells were stained with anti-PAF49 labeled with FlexAble CoraLite® Plus 488 Kit (KFA041, green), anti-Tubulin (66240-1-Ig) labeled with FlexAble CoraLite® Plus 555 Kit (KFA042, yellow), anti-Lamin labeled with FlexAble CoraLite® Plus 647 Kit (KFA043, magenta) and DAPI (blue). Confocal images were acquired with a 100x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

  • Immunofluorescence of HeLa: PFA-fixed HeLa cells were stained with anti-Lamin labeled with FlexAble CoraLite® Plus 488 Kit (KFA041, green), anti-GNL3 (67169-1-Ig) labeled with FlexAble CoraLite® Plus 555 Kit (KFA042, red), anti-Tom20 (11802-1-AP) labeled with FlexAble CoraLite® Plus 647 Kit (KFA003, magenta) and DAPI (blue). Confocal images were acquired with a 100x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

    Immunofluorescence of HeLa: PFA-fixed HeLa cells were stained with anti-Lamin labeled with FlexAble CoraLite® Plus 488 Kit (KFA041, green), anti-GNL3 (67169-1-Ig) labeled with FlexAble CoraLite® Plus 555 Kit (KFA042, red), anti-Tom20 (11802-1-AP) labeled with FlexAble CoraLite® Plus 647 Kit (KFA003, magenta) and DAPI (blue). Confocal images were acquired with a 100x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

  • Immunofluorescence of HeLa: PFA-fixed HeLa cells were stained with anti-Tubulin (66240-1-Ig) labeled with FlexAble CoraLite® Plus 488 Kit (KFA041, green), anti-Lamin labeled with FlexAble CoraLite® Plus 647 Kit (KFA043, magenta), CoraLite® 594-conjugated GNL3 antibody (CL594-67169) and DAPI (blue). Confocal images were acquired with a 100x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

    Immunofluorescence of HeLa: PFA-fixed HeLa cells were stained with anti-Tubulin (66240-1-Ig) labeled with FlexAble CoraLite® Plus 488 Kit (KFA041, green), anti-Lamin labeled with FlexAble CoraLite® Plus 647 Kit (KFA043, magenta), CoraLite® 594-conjugated GNL3 antibody (CL594-67169) and DAPI (blue). Confocal images were acquired with a 100x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

  • Immunofluorescence of HeLa: PFA-fixed HeLa cells were stained with anti-GNL3 (67169-1-Ig) labeled with FlexAble CoraLite® Plus 488 Kit (KFA041, green), mouse anti-Lamin and anti-mouse IgG secondary antibody Alexa Fluor® 568 (red), rabbit anti-COXIV (11242-1-AP) and anti-rabbit IgG secondary antibody Alexa Fluor® 647 (cyan) and DAPI (blue). Confocal images were acquired with a 100x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

    Immunofluorescence of HeLa: PFA-fixed HeLa cells were stained with anti-GNL3 (67169-1-Ig) labeled with FlexAble CoraLite® Plus 488 Kit (KFA041, green), mouse anti-Lamin and anti-mouse IgG secondary antibody Alexa Fluor® 568 (red), rabbit anti-COXIV (11242-1-AP) and anti-rabbit IgG secondary antibody Alexa Fluor® 647 (cyan) and DAPI (blue). Confocal images were acquired with a 100x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

  • Immunofluorescence of HeLa: PFA-fixed HeLa cells were stained with anti-Tubulin (66240-1-Ig) labeled with FlexAble CoraLite® Plus 488 Kit (KFA041, green). Cell nuclei are in blue. Confocal images were acquired with a 100x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

    Immunofluorescence of HeLa: PFA-fixed HeLa cells were stained with anti-Tubulin (66240-1-Ig) labeled with FlexAble CoraLite® Plus 488 Kit (KFA041, green). Cell nuclei are in blue. Confocal images were acquired with a 100x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

  • Flow cytometry of PBMC. 1X10^6 human peripheral blood mononuclear cells (PBMCs) were stained with anti-human CD45 (clone HI30, 65109-1-Ig) labeled with FlexAble CoraLite® Plus 750 Kit (KFA024) and anti-human CD3 (clone OKT3, 65133-1-Ig) labeled with FlexAble CoraLite® Plus 488 Kit (KFA041).

    Flow cytometry of PBMC. 1X10^6 human peripheral blood mononuclear cells (PBMCs) were stained with anti-human CD45 (clone HI30, 65109-1-Ig) labeled with FlexAble CoraLite® Plus 750 Kit (KFA024) and anti-human CD3 (clone OKT3, 65133-1-Ig) labeled with FlexAble CoraLite® Plus 488 Kit (KFA041).

  • Flow cytometry of PBMC. 1X10^6 human peripheral blood mononuclear cells (PBMCs) were stained with anti-human CD3 (clone UCHT1, 65151-1-Ig) labeled with FlexAble CoraLite® Plus 7505 Kit (KFA024) and anti-human CD8 (clone UCHT4, 65204-1-Ig) labeled with FlexAble CoraLite® Plus 488 Kit (KFA041).

    Flow cytometry of PBMC. 1X10^6 human peripheral blood mononuclear cells (PBMCs) were stained with anti-human CD3 (clone UCHT1, 65151-1-Ig) labeled with FlexAble CoraLite® Plus 7505 Kit (KFA024) and anti-human CD8 (clone UCHT4, 65204-1-Ig) labeled with FlexAble CoraLite® Plus 488 Kit (KFA041).

  • WB of HeLa and HepG2 cell lysates: HeLa and HepG2 cell lysates were detected with anti-HSP90 (60318-1-Ig) labeled with FlexAble CoraLite® Plus 488 Kit (KFA041, green), anti-CACYBP (68161-1-Ig) labeled with FlexAble CoraLite® Plus 555 Kit (KFA042, red) and anti-TUFM (67802-1-Ig) labeled with FlexAble CoraLite® Plus 647 Kit (KFA043, blue).

    WB of HeLa and HepG2 cell lysates: HeLa and HepG2 cell lysates were detected with anti-HSP90 (60318-1-Ig) labeled with FlexAble CoraLite® Plus 488 Kit (KFA041, green), anti-CACYBP (68161-1-Ig) labeled with FlexAble CoraLite® Plus 555 Kit (KFA042, red) and anti-TUFM (67802-1-Ig) labeled with FlexAble CoraLite® Plus 647 Kit (KFA043, blue).

产品介绍


Product name                

FlexAble CoraLite® Plus 488 Antibody Labeling Kit for Mouse IgG2a                

Assay type                

Antibody labeling                

Tested applications                

IF, FC, WB                

Species Reactivity                

Mouse IgG2a               

Antibody amount per labeling reaction                

0.5 µg antibody                

Conjugate                

CoraLite® Plus 488               

Excitation / Emission maxima wavelengths               

493 nm / 522 nm                


Component            10 rxns            50 rxns
           		4×50 rxns            
CoraLite® Plus 488 FlexLinker for Mouse IgG2a10 μL50 μL4×50 μL
FlexQuencher for Mouse IgG2a20 μL100 μL4×100 μL
FlexBuffer100 μL500 μL4×500 μL


Storage Condition

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

FAQ

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. 


More FAQs