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Purified Mouse Anti-p150 [Glued]
Purified Mouse Anti-p150 [Glued]

Western blot analysis of p150 [Glued] on a rat cerebrum lysate (left). Lane 1: 1:1000, lane 2: 1: 2000, lane 3: 1:4000 dilution of the mouse anti- p150 [Glued] antibody.

Immunofluorescent staining of SK-N-SH cells (Human neuroblastoma; ATCC HTB-11) (right). Cells were seeded in collagen coated 384-well imaging microplates (Material # 353962) at ~ 8,000 cells per well.  After overnight incubation, cells were stained using the Triton-X 100 fix/perm protocol (see Recommended Assay Procedure) with the mouse anti- p150 [Glued] antibody and counter-stained with Hoechst 33342 (pseudo-colored blue).  The second step reagent used was Alexa Fluor® 488 goat anti-mouse Ig (pseudo-colored green) (Invitrogen).  The images were captured on a BD Pathway™ 855 or 435 Bioimager system using a 20x objective and merged using BD Attovision™ software.  This antibody also stained SH-SY5Y (Human neuroblastoma; ATCC CRL-2266)  and C6 (Rat glioma; ATCC CCL-107) cells using both the Triton-X 100 and methanol fix/perm protocols (see Recommended Assay Procedure).

Western blot analysis of p150 [Glued] on a rat cerebrum lysate (left). Lane 1: 1:1000, lane 2: 1: 2000, lane 3: 1:4000 dilution of the mouse anti- p150 [Glued] antibody.

Immunofluorescent staining of SK-N-SH cells (Human neuroblastoma; ATCC HTB-11) (right). Cells were seeded in collagen coated 384-well imaging microplates (Material # 353962) at ~ 8,000 cells per well.  After overnight incubation, cells were stained using the Triton-X 100 fix/perm protocol (see Recommended Assay Procedure) with the mouse anti- p150 [Glued] antibody and counter-stained with Hoechst 33342 (pseudo-colored blue).  The second step reagent used was Alexa Fluor® 488 goat anti-mouse Ig (pseudo-colored green) (Invitrogen).  The images were captured on a BD Pathway™ 855 or 435 Bioimager system using a 20x objective and merged using BD Attovision™ software.  This antibody also stained SH-SY5Y (Human neuroblastoma; ATCC CRL-2266)  and C6 (Rat glioma; ATCC CCL-107) cells using both the Triton-X 100 and methanol fix/perm protocols (see Recommended Assay Procedure).

Product Details
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BD Transduction Laboratories™
Rat (QC Testing), Human, Mouse, Dog (Tested in Development)
Mouse IgG1
Rat p150 [Glued] aa. 3-202
Bioimaging, Western blot (Routinely Tested), Immunofluorescence (Tested During Development)
150 kDa
250 µg/ml
AB_399948
Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at -20°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

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

  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.
612709 Rev. 1
Antibody Details
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12/P150GLUED

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.

612709 Rev. 1
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
612709 Rev.1
Citations & References
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Development References (2)

  1. 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
  2. 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
612709 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.