![Purified Mouse Anti-p150 [Glued]](/content/dam/bdb/products/global/reagents/microscopy-imaging-reagents/immunofluorescence-reagents/612xxx/6127xx/612708_base/612708Image1.png)
Preparation And Storage
Recommended Assay Procedures
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.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
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.
Development References (2)
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Holzbaur EL, Hammarback JA, Paschal BM, Kravit NG, Pfister KK, Vallee RB. Homology of a 150K cytoplasmic dynein-associated polypeptide with the Drosophila gene Glued. 1991; 351(6327):579-583. (Biology). View Reference
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Waterman-Storer CM, Karki S, Holzbaur EL. The p150Glued component of the dynactin complex binds to both microtubules and the actin-related protein centractin (Arp-1). 1995; 92(5):1634-1638. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.