Sialylated Mucin-Type-O-Glycans Antibody (clone FW17)

LifeSpan BioSciences

Sialylated Mucin-Type-O-Glycans Antibody (clone FW17)

23

Sialylated Mucin-Type-O-Glycans.

Tumor and normal tissues were first fixed in 6% neutral phosphate-buffered formalin and dehydrated in ethanol by standard histological techniques. After embedding all tissues in paraffin, serial microtome tissue sections (5 microm) were used for comparative immunohistological studies. Reactivity of hybridoma tissue culture supernatants was determined using a four-step immunoperoxidase technique. After deparaffinization in xylene, rehydration through graded ethanol, and blocking endogenous peroxidase activity with 2% H202 in absolute methanol for 30 min, the sections were incubated as follows: (1) normal swine serum (5% in PBS), (2) MAb in tissue culture supernatant or mouse ascites fluid diluted to 1/100 in PBS, (3) rabbit anti-mouse immunoglobulin (2% in PBS), (4) swine anti-rabbit immunoglobulin (2% in PBS), (5) rabbit peroxidase anti-peroxidase (PAP) complex. The slides were treated with 0.2 mg/ml - 3-amino-9-ethylcarbazole in 0.02 M sodium acetate buffer containing 0.01% H202, then counterstained with hematoxylin, and mounted in glycerol jelly. All incubation steps were performed in a moist chamber at 21°C for 30 min.. ELISA, ICC, IHC, IHC-P Tumor and normal tissues were first fixed in 6% neutral phosphate-buffered formalin and dehydrated in ethanol by standard histological techniques. After embedding all tissues in paraffin, serial microtome tissue sections (5 µm) were used for comparative immunohistological studies. Reactivity of hybridoma tissue culture supernatants was determined using a four-step immunoperoxidase technique. After deparaffinization in xylene, rehydration through graded ethanol, and blocking endogenous peroxidase activity with 2% H202 in absolute methanol for 30 min, the sections were incubated as follows: (1) normal swine serum (5% in PBS), (2) MAb in tissue culture supernatant or mouse ascites fluid diluted to 1/100 in PBS, (3) rabbit anti-mouse immunoglobulin (2% in PBS), (4) swine anti-rabbit immunoglobulin (2% in PBS), (5) rabbit peroxidase anti-peroxidase (PAP) complex. The slides were treated with 0.2 mg/ml - 3-amino-9-ethylcarbazole in 0.02 M sodium acetate buffer containing 0.01% H202, then counterstained with hematoxylin, and mounted in glycerol jelly. All incubation steps were performed in a moist chamber at 21°C for 30 min.

ELISA, ImmunoCytoChemistry, ImmunoHistoChemistry, ImmunoHistoChemistry Paraffin

Research Use Only

Monoclonal

FW17

Unconjugated

...

Mouse

IgM

Human

-20°C

12352203