MAXbind™ Staining Medium
Catalog No.: 15253
Format: 250 ml
Applications: Immunofluorescence, Immunohistochemistry, Western blotting & ELISAFormulation: Non-mammalian antibody binding agent in PBS, pH 7.4, containing 0.1% Triton X-100 and 0.09% sodium azide
Storage: Store at 4°C. Guaranteed stable for 6 months when stored properly.
Concentration: MAXbind is to be used at the delivered concentration (1X).
Description: MAXbind Staining Medium is a non-mammalian incubation agent that optimizes antibody staining in immunofluorescenceand immunostaining assays. Used in combination with MAXblock™ Blocking Medium (Cat. No. 15252), MAXbind promotes superior
antibody binding while helping to eliminate non-specific primary and secondary antibody binding.
Quality Control: MAXbind was tested for effectiveness in promoting antibody binding in immunofluorescence (below) as well as in IHC
Instructions: For immunofluorescence, immunohistochemistry or immunocytochemistry, cells can be grown either directly on slides orcoverslips, or spun down onto slides or coverslips. The volumes below assume cells grown on coverslips in the wells of a 6-well plate.
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After blocking cells with MAXblock or similar for 1 hour in a humidified environment, e.g. an incubator or slide warmer, aspirate
the MAXblock and add 1 ml 1X MAXwash™ Washing Medium (Cat. No. 15254), or similar. Rock the plate for 10 minutes on a rotatingplatform. During this wash step, dilute your primary antibody to an appropriate dilution in MAXbind. For a coverslip in the well of a
6-well plate, you will need to add 1 ml. To make a 1:500 dilution, dilute 2 µl primary antibody in 1 ml MAXbind per well.
Aspirate the MAXwash and add 1 ml of the diluted primary antibody to each well. Incubate for 1 hour at 37°C in a humid environment, e.g. an incubator or slide warmer.
Aspirate the diluted primary antibody from the cells, add 1 ml 1X MAXwash and rock the plate for 10 minutes on a rotating platform.
Aspirate the MAXwash and repeat 2 more times for a total of 3 washes. During the last wash, dilute your secondary antibody in
MAXbind to a dilution of 1:500 to 1:2000. You will need to add 1 ml of diluted secondary antibody per well.
Incubate the diluted secondary antibody for 1 hour at 37°C in a darkened, humid environment, e.g. an incubator or slide warmer.
From here forward, it is important to limit the amount of light exposure to the fluorescent dye on the secondary antibody.
Aspirate the diluted secondary antibody from the cells and add 1 ml 1X MAXwash. Rock the plate for 10 minutes on a rotating
platform. Aspirate the MAXwash and repeat 3 more times for a total of 4 washes.
After the last wash, carefully remove the coverslip from the well using flat-edged forceps. In some cases, the coverslip is slightly
stuck to the bottom of the well and may need to be dislodged using a beveled needle. Using the needle, carefully stand up thecoverslip in the well and grab it with forceps.
Dry the coverslip to remove excess MAXwash. Hold the corner of the coverslip to a Kimwipe to remove any excess MAXwash fromthe coverslip. Remember to limit light exposure. You are now ready for counterstaining and slide mounting.

MAXbind is for in vitro research use only and is not intended for use in humans or animals.
Immunofluorescence staining with MAXbind.
Bovine pulmonary artery endothelial cells were labeled for
mitochondria (red), F-actin (green) and DAPI (blue) using
MAX Stain™ immunofluorescence and immunostaining
products. In addition to MAXbind, MAXblock, MAXwash andMAXfluor™ DAPI were used in the creation of this image.

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