M176-A48
Lot 003~
Page 1For Research Use Only.
Not for use in diagnostic procedures.

Anti-EEA1 mAb-Alexa Fluor® 488
CODE No.

M176-A48

CLONALITY
CLONEISOTYPE
QUANTITY

Monoclonal
3C10
Mouse IgG2a κ
100 µL, 1 mg/mL

SOURCEIMMUNOGEN
FORMULATIONSTORAGE

Purified IgG from hybridoma supernatant
Human EEA1, N-terminal (synthetic peptide)
PBS containing 1% BSA and 0.1% ProClin150.
This antibody solution is stable for one year from the date of purchase when stored at 4°C.

APPLICATION-CONFIRMED
Immunocytochemistry

10 µg/mL

SPECIES CROSS REACTIVITY on WB
Species

HumanMouse

Rat

Hamster

Cells

HeLa, A549

NIH/3T3, MEF

NRKNot tested

Reactivity+++Entrez Gene ID

8411 (Human), 216238 (Mouse), 314764 (Rat)

REFERENCES

1) Gaullier, J. M., et al., J. Biol. Chem. 275, 24595-24600 (2000)
2) Mu, F. T., et al., J. Biol. Chem. 270, 13503-13511 (1995)

For more information, please visit our web site https://ruo.mbl.co.jp/

LABEL LICENSES:This product is provided under an agreement between LIFE TECHNOLOGIES Corporation, and Medical & Biological Laboratories Co.,LTD, and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications, andcorresponding non-U.S. equivalents, owned by or licensed to LIFE TECHNOLOGIES Corporation. For information on purchasing a license
to this product for any other use, contact LIFE TECHNOLOGIES Corporation, Molecular Probes Detection Technologies, BusinessDevelopment, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

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Lot 003~Page 2
The descriptions of the following protocols are examples. Each user should determine the appropriate condition.

Immunocytochemistry1) Spread the cells in the nutrient condition on a glass slide, then incubate in a CO2 incubator for one night.
2) Remove the culture supernatant by careful aspiration.
3) Fix the cells by immersing the slide in 4% paraformaldehyde (PFA)/PBS for 10 min. at room temperature (20~25°C).
4) Prepare a wash container such as a 500 mL beaker with a magnetic stirrer. Then wash the fixed cells on the glass slide bysoaking the slide with a plenty of PBS in the wash container for 5 min. Take care not to touch the cells. Repeat another wash
twice more.
5) Immerse the slide in 0.2% Triton X-100/PBS for 10 min. at room temperature.
6) Wash the slide 2 times with PBS.
7) Add 200 µL of the primary antibody diluted with 2% fetal calf serum (FCS)/PBS as suggested in the APPLICATIONS onto
the cells and incubate for 30 min. at room temperature. (Optimization of antibody concentration or incubation condition isrecommended if necessary.)8) Wash the slide 2 times with PBS.
9) Counter stain with DAPI for 5 min. at room temperature.
10) Wash the slide 2 times with PBS.
11) Wipe excess liquid from slide but take care not to touch the cells. Never leave the cells to dry.
12) Promptly add mounting medium onto the slide, then put a cover slip on it.
(Positive control for Immunocytechemistry; HeLa)

Immunocytochemical detection of EEA1 in HeLa
Green: M176-A48
Blue: DAPI210316-3