Description
p150 [Glued] was identified as a polypeptide associated with cytoplasmic dynein,the minus-end-directed microtubule-based motor protein. p150 [Glued] is also a member of the oligomeric dynactin complex. Dynactin mediates dynein-driven vesicle motility, as well as lower eukaryote nuclear transport. p150 [Glued] bears significant homology to the product of the Glued gene in Drosophila. It has been shown in vitro to be a required activator of dynein-mediated transport along microtubules. The p150 [Glued] component of the dynactin complex binds to microtubules and the actin-like protein Centractin (Arp-1), another member of the dynactin complex. In the developing rat, p150 [Glued] is expressed at high levels in neural tissue. Microtubule binding assays with selected constructs of p150 [Glued] indicate that amino acids 39-150 are required for microtubule association.
Western blot: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml Bioimaging: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Bioimaging_Certified.shtml Methanol Procedure for a 96 well plate: Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 90% methanol. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Wash three times with PBS. Flick out PBS and add second step reagent. Incubate for 1 hour at RT. Wash three times with PBS. Image sample. Triton-X 100 Procedure for a 96 well plate: Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 0.1% Triton-X 100. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Flick out and wash three times with PBS. Flick out and add second step reagent. Incubate for 1 hour at RT. Flick out and wash three times with PBS. Image sample.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at -20°C.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
