Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of MCF7 (Lane 1), A-431 (Lane 2), H1975 (Lane 3), PC-3 (Lane 4), DU-145 (Lane 5), MDA-MB-231 (Lane 6), Ramos (Lane 7), HEK-293 (Lane 8), 3T3-L1 (Lane 9), Jurkat (Lane 10) and A549 (Lane 11). The blots were probed with Rabbit Anti-GRB2 Polyclonal Antibody (Cat. No. PA5-17692, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Cat. No. A27036, 0.25 µg/mL, 1:4000 dilution). A 25 kDa band corresponding to GRB2 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex NuPAGE 12% Bis-Tris gel (Cat. No. NP0342BOX), XCell SureLock Electrophoresis System (Cat. No. EI0002) and Novex Sharp Pre-Stained Protein Standard (Cat. No. LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane using iBlot 2 Dry Blotting System (Cat. No. IB21001). The membrane was probed with the relevant primary and secondary antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce ECL Western Blotting Substrate (Cat. No. 32106).
The plasma membrane is a selectively permeable phospholipid bilayer that surrounds the cytoplasm of cells, separating the intracellular components from the extracellular environment. It is primarily composed of proteins and lipids and is instrumental in signaling pathways. The plasma membrane also anchors the cytoskeleton, giving the cell shape, and serves as an attachment point for the extracellular matrix and surrounding cells to help them group together to form tissues.
Plasma membrane marker antibodies detect proteins specific to the plasma membrane and aid in the study of the morphology and functions of the plasma membrane. Plasma membrane marker antibodies can also help elucidate the role a protein may play in tasks that are centered in the plasma membrane such as transporter, receptor, anchor, enzyme, etc.
Invitrogen plasma membrane marker antibodies are designed to dependably detect the key plasma membrane targets. Each antibody is validated for use in various applications. Key plasma membrane targets include:
Plasma membrane antibody targets
Applications
Each Invitrogen plasma membrane marker antibody is validated for use in applications such as ELISA, western blot, flow cytometry, immunofluorescence, immunohistochemistry, immunocytochemistry, and immunoprecipitation.
Immunofluorescent detection of Zo-1 in MDCK cells. Confluent monolayers were fixed in 50% methanol/50% Acetone, blocked for at least 30 minutes in 1% BSA then incubated 2 hours with a Zo-1 monoclonal antibody (Cat. No. 33-9100) at 5 µg/mL, washed, then incubated 1 hour with Alexa Fluor 488 conjugated Donkey anti-Mouse secondary antibody (Cat. No. A-21202) at a dilution of 1:2000. Cells were counterstained with DAPI (blue). Coverslips were mounted with Prolong Gold Antifade reagent (Cat. No. P36930) and imaged at 40X. Images generated by Joell Solan in Paul Lampe Lab at the Fred Hutchinson cancer Research Center.
Resources
Downloads
Product selection guides
Learning and support
For Research Use Only. Not for use in diagnostic procedures.
For Research Use Only. Not for use in diagnostic procedures.