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FlexAble Biotin Antibody Labeling Kit for Rabbit IgG

Application

IF, FC, IHC

Reactivity

Rabbit

Conjugate

Biotin

Cat No : KFA007

Print Protocol

Validation Data Gallery

  • Immunofluorescence of HeLa: PFA-fixed HeLa cells were stained with anti-GM130 (11308-1-AP) labeled with FlexAble Biotin Kit for Rabbit IgG (KFA007) and Streptavidin-ATTO594​ (yellow). Nuclei are stained with 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-GM130 (11308-1-AP) labeled with FlexAble Biotin Kit for Rabbit IgG (KFA007) and Streptavidin-ATTO594​ (yellow). Nuclei are stained with 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 B1 antibody (12987-1-AP) followed by polyclonal goat anti-rabbit AF488 secondary antibody (green), anti-GM130 antibody (11308-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit (KFA007) and Streptavidin-ATTO594​ (yellow), and anti-Tom20 antibody (11802-1-AP) separately labeled with FlexAble Biotin Antibody Labeling Kit (KFA007) and Streptavidin-AQ650 (red). Nuclei are stained with 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 B1 antibody (12987-1-AP) followed by polyclonal goat anti-rabbit AF488 secondary antibody (green), anti-GM130 antibody (11308-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit (KFA007) and Streptavidin-ATTO594​ (yellow), and anti-Tom20 antibody (11802-1-AP) separately labeled with FlexAble Biotin Antibody Labeling Kit (KFA007) and Streptavidin-AQ650 (red). Nuclei are stained with 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 mouse testis: FFPE mouse testis sections were stained with anti-TNP1 antibody (17178-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit for Rabbit IgG (KFA007) and Streptavidin-488 (green), and anti-BOLL antibody (13720-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit for Rabbit IgG (KFA007) and Streptavidin-594 (red).  Cell nuclei are stained with DAPI (blue). 

    Immunofluorescence of mouse testis: FFPE mouse testis sections were stained with anti-TNP1 antibody (17178-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit for Rabbit IgG (KFA007) and Streptavidin-488 (green), and anti-BOLL antibody (13720-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit for Rabbit IgG (KFA007) and Streptavidin-594 (red).  Cell nuclei are stained with DAPI (blue). 

  • Flow cytometry of PBMC.  1X10^6 HEK293 cells were fixed with FoxP3/Transcription Factor Staining Buffer Kit (PF00011), then stained with anti-human Mitofilin antibody (10179-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit for Rabbit IgG (KFA007) and Streptavidin-APC. Single cells are gated.

    Flow cytometry of PBMC.  1X10^6 HEK293 cells were fixed with FoxP3/Transcription Factor Staining Buffer Kit (PF00011), then stained with anti-human Mitofilin antibody (10179-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit for Rabbit IgG (KFA007) and Streptavidin-APC. Single cells are gated.

  • Flow cytometry of PBMC.  1X10^6 HEK293 cells were fixed with FoxP3/Transcription Factor Staining Buffer Kit (PF00011), then stained with anti-human Mitofilin antibody (10179-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit for Rabbit IgG (KFA007) and Streptavidin-PE. Single cells are gated.

    Flow cytometry of PBMC.  1X10^6 HEK293 cells were fixed with FoxP3/Transcription Factor Staining Buffer Kit (PF00011), then stained with anti-human Mitofilin antibody (10179-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit for Rabbit IgG (KFA007) and Streptavidin-PE. Single cells are gated.

  • Immunohistochemical analysis of paraffin-embedded Rabbit colon tissue using anti-Cytokeratin 20 antibody (17329-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit for Rabbit IgG (KFA007) and used at a dilution of 1:1000 (under 10x lens). Streptavidin Poly-HRP and DAB substrate was used for detection. Heat mediated antigen retrieval performed with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded Rabbit colon tissue using anti-Cytokeratin 20 antibody (17329-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit for Rabbit IgG (KFA007) and used at a dilution of 1:1000 (under 10x lens). Streptavidin Poly-HRP and DAB substrate was used for detection. Heat mediated antigen retrieval performed with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded Rabbit colon tissue using anti-Cytokeratin 20 antibody (17329-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit for Rabbit IgG (KFA007) and used at a dilution of 1:1000 (under 40x lens). Streptavidin Poly-HRP and DAB substrate was used for detection. Heat mediated antigen retrieval performed with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded Rabbit colon tissue using anti-Cytokeratin 20 antibody (17329-1-AP) labeled with FlexAble Biotin Antibody Labeling Kit for Rabbit IgG (KFA007) and used at a dilution of 1:1000 (under 40x lens). Streptavidin Poly-HRP and DAB substrate was used for detection. Heat mediated antigen retrieval performed with Tris-EDTA buffer (pH 9.0).

产品介绍


Product name                

FlexAble Biotin Antibody Labeling Kit for Rabbit IgG                

Assay type                

Antibody labeling                

Tested applications                

IF, FC, IHC               

Species Reactivity                

Rabbit IgG                

Antibody amount per labeling reaction            

0.5 µg antibody                               

Conjugate                

Biotin               

Compatible with               

Streptavidin conjugated dyes and molecules 



Component            10 rxns            50 rxns
           		4×50 rxns            
Biotin FlexLinker for Rabbit IgG10 μL50 μL4×50 μL
FlexQuencher for Rabbit IgG20 μL100 μL4×100 μL
FlexBuffer100 μL500 μL4×500 μL


保存条件

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

其他

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