FlexAble Biotin Antibody Labeling Kit for Mouse IgG1

• 

  Immunofluorescence of HeLa: PFA-fixed HeLa cells were stained with anti-CD147 antibody labeled with FlexAble Biotin Antibody Labeling Kit for Mouse IgG1 (KFA027) 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.

• 

  Flow Cyotometry of PBMC.  1X10^6 human PBMC were stained with anti-human CD86 antibody (65156-1-Ig) labeled with FlexAble Biotin Antibody Labeling Kit for Mouse IgG1 (KFA027) and Streptavidin-APC. The cells were co-stained with either mouse IgG2b isotype control or anti-human CD14 antibody (CL488-65246).  Cells are not fixed, Monocytes are gated. 

• 

  Flow cytometry of PBMC. 1X10^6 human PBMC were stained with anti-human CD86 antibody (65156-1-Ig) labeled with FlexAble Biotin Antibody Labeling Kit for Mouse IgG1 (KFA027) and Streptavidin-PE.  The cells were co-stained with either mouse IgG2b isotype control or anti-human CD14 antibody (CL488-65246).  Cells are not fixed, Monocytes are gated.

• 

  Flow cytometry of PBMC. 1X10^6 human PBMC were stained with anti-human CD86 antibody (65156-1-Ig) labeled with FlexAble Biotin Antibody Labeling Kit for Mouse IgG1 (KFA027) and Streptavidin-PE.  The cells were co-stained with either mouse IgG2b isotype control or anti-human CD14 antibody (CL488-65246).  Cells are not fixed, Monocytes are gated.

• 

  Immunohistochemical analysis of paraffin-embedded human oesophagus cancer tissue slide using anti-Cytokeratin 5 antibody (66727-1-Ig) labeled with FlexAble Biotin Antibody Labeling Kit for Mouse IgG1 (KFA027) and used at a dilution of 1:400 (under 10x lens & 40xlens). 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 human oesophagus cancer tissue slide using anti-Cytokeratin 5 antibody (66727-1-Ig) labeled with FlexAble Biotin Antibody Labeling Kit for Mouse IgG1 (KFA027) and used at a dilution of 1:400 (under 10x lens & 40xlens). Streptavidin Poly-HRP and DAB substrate was used for detection. Heat mediated antigen retrieval performed with Tris-EDTA buffer (pH 9.0).

• 

  Flow Cyotometry of PBMC.  1X10^6 human PBMC were stained with anti-human CD86 antibody (65156-1-Ig) labeled with FlexAble Biotin Antibody Labeling Kit for Mouse IgG1 (KFA027) and Streptavidin-APC. The cells were co-stained with either mouse IgG2b isotype control or anti-human CD14 antibody (CL488-65246).  Cells are not fixed, Monocytes are gated. 

Cat No. KFA027

Applications

IF, FC, IHC

Reactivity

Mouse

Conjugate

Biotin

---

Product Information

Product name	FlexAble Biotin Antibody Labeling Kit for Mouse IgG1
Assay type	Antibody labeling
Tested applications	IF, FC, IHC
Species Reactivity	Mouse IgG1
Antibody amount per labeling reaction	0.5 µg antibody
Conjugate	Biotin
Compatible with	Streptavidin conjugated dyes and molecules

Kit Components

Component	10 rxns	50 rxns	200rxns
FlexAble Biotin Antibody Labeling Kit for Mouse IgG1			50rxns
FlexBuffer	100ul	500ul
FlexLinker for Mouse IgG1 Biotin	10ul	50ul
FlexQuencher for Mouse IgG1	20ul	100ul

Storage Condition

Store for 1 year at -20°C or for 6 months at +4°C upon receipt. 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