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Purified Mouse Anti-β-Tubulin 5H1 RUO

Purified Mouse Anti-β-Tubulin 

Clone  5H1   (RUO)

  • Brand BD Pharmingen™
  • Concentration 0.5 mg/ml
  • Isotype Mouse IgM, κ
  • Reactivity Human (QC Testing)  Mouse, Rat, Cow (Tested in Development) 
  • Application Western blot (Routinely Tested) 
    Bioimaging, Immunohistochemistry (Tested During Development) 
  • Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
  • Regulatory Status RUO
Product Details Recommended Assay References

Description

Tubulin is a highly conserved protein with a molecular weight of ~50 kD. The self-assembly of tubulin leads to microtubules, hollow cylinders that are one of the major components of the eukaryotic cytoskeleton. Microtubules play key roles in chromosome segregation in mitosis, intracellular transport, ciliary and flagellar bending, and structural support of the cytoskeleton. There are two main classes of tubulin isoforms, α- and β-tubulin, which are usually products of separate genes. Microtubules are made from protofilaments, strings of alternating α- and β-tubulin spaced 4 nm apart and pointing in the same direction. Tubulin can be posttranslationally modified in several ways, including phosphorylation, acetylation, glutamylation, and detyrosination. For example, microtubules that turn over slowly tend to be acetylated and detyrosinated.

The 5H1 monoclonal antibody reacts with β-tubulin. It does not cross-react with α-tubulin.

Format
  • Format Purified

Other Formats →

Suggested Companion Products

Fixation Buffer RUO
100 mL Cat No: 554655

Perm Buffer III RUO
125 mL Cat No: 558050

Stain Buffer (FBS) RUO
500 mL Cat No: 554656

FITC Goat Anti-Mouse Ig Polyclonal RUO
0.5 mg Cat No: 554001

HRP Goat Anti-Mouse Ig RUO
1 mL Cat No: 554002

Resources & Tools
 Spectrum Viewer  Download TDS  Regulatory Document Website
Preparation and Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
  3. Triton is a trademark of the Dow Chemical Company.
  4. 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.
  5. Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.


Bioimaging

1. Seed the cells in appropriate culture medium at ~10,000 cells per well in a BD Falcon™ 96-well Imaging Plate (Cat. No. 353219) and culture overnight.

2. Remove the culture medium from the wells, and fix the cells by adding 100 μl of BD Cytofix™ Fixation Buffer (Cat. No. 554655) to each well. Incubate for 10 minutes at room temperature (RT).

3. Remove the fixative from the wells, and permeabilize the cells using either BD Perm Buffer III, 90% methanol, or Triton™ X-100:

a. Add 100 μl of -20°C 90% methanol or Perm Buffer III (Cat. No. 558050) to each well and incubate for 5 minutes at RT.

OR

b. Add 100 μl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.

4. Remove the permeabilization buffer, and wash the wells twice with 100 μl of 1× PBS.

5. Remove the PBS, and block the cells by adding 100 μl of BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) to each well. Incubate for 30 minutes at RT.

6. Remove the blocking buffer and add 50 μl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.

7. Remove the primary antibody, and wash the wells three times with 100 μl of 1× PBS.

8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 μl to each well, and incubate in the dark for 1 hour at RT.

9. Remove the second step reagent, and wash the wells three times with 100 μl of 1× PBS.

10. Remove the PBS, and counter-stain the nuclei by adding 200 μl per well of 2 μg/ml Hoechst 33342 (e.g., Sigma-Aldrich Cat. No. B2261) in 1× PBS to each well at least 15 minutes before imaging.

11. View and analyze the cells on an appropriate imaging instrument.

Bioimaging: For more detailed information please refer to http://www.bdbiosciences.com/support/resources/protocols/ceritifed_reagents.jsp

Western blot: For more detailed information please refer to http://www.bdbiosciences.com/support/resources/protocols/monoclonal_anti.jsp


  1. Brown KD, Wood KW, Cleveland DW. The kinesin-like protein CENP-E is kinetochore-associated throughout poleward chromosome segregation during anaphase-A. J Cell Sci. 1996; 109:961-969.

  2. Cho J-H, Johnson GVW. Primed phosphorylation of tau at Thr231 by glycogen synthase kinase 3β (GSK3β) plays a critical role in regulating tau's ability to bind and stabilize microtubules. J Neurochem. 2004; 88:349-358.

  3. Helfand BT, Mikami A, Vallee RB, Goldman RD. A requirement for cytoplasmic dynein and dynactin in intermediate filament network assembly and organization. J Cell Biol. 2002; 157(5):795-806.

  4. Prahlad V, Yoon M, Moir RD, Vale RD, Goldman RD. Rapid movements of vimentin on microtubule tracks: kinesin-dependent assembly of intermediate filament networks. J Cell Biol. 1998; 143(1):159-170.

  5. Wang Y, Loomis PA, Zinkoswski RP, Binder LI. A novel tau transcript in cultured human neuroblastoma cells expressing nuclear tau. J Cell Biol. 1993; 121(2):257-267.

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