Western Blotting (WB), Immunohistochemistry (IHC), Flow Cytometry (FACS)
Specificity
JSB-1 reacts with a conserved cytoplasmic epitope of the human plasma membrane- associated 170-180 kDa glycoprotein, the expression of which is strongly correlated with the degree of multidrug resistance (MDR) derived MDR cell lines and human MDR cell lines, including cell lines derived from lung, ovaries and B cell lymphomas.
Flow cytometry: Use at least a 1:10 dilution. Fix cells in 10% (v/v) Lysing Solution, followed by primary antibody and anti-mouse- FITC. Immunocytochemistry: Use at least a 1:20 dilution. (acetone-fixed cell preparation) Immunohistochemistry: acetone-fixed frozen sections or formalin-fixed, paraffin-embedded tissues: Use at a 1:20 dilution. Optimal staining results are obtained with routine 2-step ABC or APAAP methods using acetone-fixed cytocentrifuge preparations or cryostat sections. In case the B-5 fixative is used (see below), paraffin-embedded tissue can also be stained with the antibody. Western blot: Start optimizing working dilution at 1:10, followed by incubation with anti- mouse HRP. B-5 Fixative Procedure: Fix tissue in freshly prepared B-5 fixative: add 2 mL of 40% formalin to 20 mL of B-5 Stock Solution. Sections must be dezenkerized to remove mercuric pigment before immunostaining. Dezenkerize by placing slides with sections in Gram's Iodine solution for 5 min. at room temperature. Wash well in distilled water. Place slides in Sodium Thiosulphate solution for 5 min. at room temperature. Wash in running tap water for 3 min. Place in PBS for further immunochemical procedures. B-5 Stock Solution: 12 g mercuric chloride 2.5 g sodium acetate Dissolve in 200 mL distilled water. Gram's Iodine: 1 g I2 2 g KI Dissolve in 300 mL distilled water. KAMIYA BIOMEDICAL COMPANY Rev. 021507 PRODUCT DATA SHEET KAMIYA BIOMEDICAL COMPANY, Seattle, WA USA Tel: (206) 575-8068 Fax: (206) 575-8094 www.k-assay.com LifeScience@k-assay.com
Restrictions
For Research Use only
Format
Liquid
Buffer
1 mL (200 tests) or 0.5 mL (100 tests) monoclonal antibody at ~250 µg IgG/mL in tissue culture supernatant with protein stabilizer and 0.1% sodium azide. Filtered through a 0.22 µm filter. Culture medium: RPMI-1640, supplemented with Nutridoma-SR. The medium does not contain serum or added enzymes.
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.