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Normal rabbit igg isotype control

Manufactured by R&D Systems
Sourced in Japan

Normal rabbit IgG isotype control is a non-specific antibody that is used as a control in immunoassays and other applications where the use of an isotype-matched control is required. It is derived from the serum of normal rabbits and does not recognize any known antigenic target.

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2 protocols using normal rabbit igg isotype control

1

Quantifying Surface Receptor Expression

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For quantification of surface receptor expression, cells were trypsinized and brought to single-cell suspension in FACS buffer (1× PBS + 2% FBS). Primary antibody was added at 1 μg/million cells in 100 μL total solution (for mouse cells: rat anti-mouse αv integrin (BD Pharmingen 551380) or rat IgG isotype control (Invitrogen); for human cells: mouse anti-αvβ3 integrin, direct PE conjugate (BioLegend 304406) or mouse IgG κ chain isotype control, direct PE conjugate (BioLegend 400112); for neuropilin-1 staining in all cells, rabbit anti-NRP-1 (Novus Biologicals NBP1-40666) or normal rabbit IgG isotype control (R&D)) and incubated for one hour on ice. For direct fluorophore-conjugated primary antibodies, cells were washed with PBS and resuspended in FACS buffer. Otherwise, after washing the cells 2× in PBS, cells were incubated with secondary fluorescently-tagged antibody (Invitrogen) for 45 minutes and washed 1× in PBS. Cells were analyzed on BD LSR-II or Fortessa HTS flow cytometers. Data were analyzed in FlowJo (TreeStar Software).
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2

Immunocytometry and Western Blotting Antibodies

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The following antibodies (Abs) were used in immunocytometry and immunohistochemistry: rabbit anti-von Willebrand factor (Factor VIII) (1:200, DAKO, Carpinterria, CA, USA), mouse anti-CD31 Ab (1:200, Abcam, Tokyo, Japan), mouse anti-Vimentin (1:200, Dako, CA, USA), mouse anti-Desmin (1:100, DAKO, Carpinterria, CA, USA), rabbit anti-hepatocyte growth factor (HGF) Ab (1:200, Abcam, Tokyo, Japan), rabbit anti-Met Ab (1:200, Abcam, Tokyo, Japan), normal Rabbit IgG isotype control (1:200, R&D Systems, Minneapolis, MN, USA), goat anti-rabbit IgG conjugated with Alexa-488 fluorescent dye (1:200, Thermofisher Scientific, Tokyo, Japan) and goat anti-mouse IgG conjugated with Alexa-594 fluorescent dye (1:200, Thermofisher Scientific, Tokyo, Japan). These antibodies were dissolved in PBS-T.
The following Abs were used in Western blotting: rabbit anti-βactin (1:1000, Biolegend, Tokyo, Japan), rabbit anti-HGF (1:1000, Abcam, Tokyo, Japan) and Goat anti-Rabbit IgG HRP conjugate (1:1000, Thermoscientific, Tokyo, Japan). These antibodies were dissolved with blocking solution (5% non-fat dried milk in PBS containing 0.5% Tween20).
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