BUB3 antibody (AA 4-16)

Catalog No. ABIN968647

Product Details anti-BUB3 Antibody

Target: BUB3 (Budding Uninhibited By Benzimidazoles 3 Homolog (Yeast) (BUB3))

Binding Specificity: AA 4-16

Reactivity: Human, Mouse, Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: This BUB3 antibody is un-conjugated

Application: Western Blotting (WB), BioImaging (BI)

Cross-Reactivity: Mouse (Murine), Rat (Rattus)

Characteristics: 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
5. 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.
6. Please refer to us for technical protocols.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human Bub3 aa. 4-16

Clone: 31-Bub3

Isotype: IgG1

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl 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 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl 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 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl 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 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN967389

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 250 μg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20°C.

References for anti-BUB3 antibody (ABIN968647)

Martinez-Exposito, Kaplan, Copeland, Sorger: "Retention of the BUB3 checkpoint protein on lagging chromosomes." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 96, Issue 15, pp. 8493-8, (1999) (PubMed).

Taylor, Ha, McKeon: "The human homologue of Bub3 is required for kinetochore localization of Bub1 and a Mad3/Bub1-related protein kinase." in: The Journal of cell biology, Vol. 142, Issue 1, pp. 1-11, (1998) (PubMed).

Target Details for BUB3

Target: BUB3 (Budding Uninhibited By Benzimidazoles 3 Homolog (Yeast) (BUB3))

Alternative Name: Bub3 (BUB3 Products)

Synonyms: BUB3L antibody, hBUB3 antibody, Aa2-050 antibody, xbub3 antibody, AU019800 antibody, AU021329 antibody, AU043350 antibody, AW146323 antibody, C78067 antibody, BUB3, mitotic checkpoint protein antibody, BUB3 mitotic checkpoint protein antibody, BUB3 mitotic checkpoint protein L homeolog antibody, BUB3 antibody, Bub3 antibody, bub3.L antibody

Background: Accurate chromosome segregation requires that all pairs of sister chromatids become appropriately attached to mitotic spindles before the onset of anaphase. Cell cycle checkpoints monitor kinetochore-microtubule interactions, so that cell cycle progression can be delayed until proper chromosome attachments are formed. In yeast, Bub1-3 genes are required for proper mitotic delay in response to unattached kinetochores. In mammals, the homologues to yeast Bub1 and Bub3 form a complex that binds kinetochores and has protein kinase activity. Bub3 contains four WD repeats, three in the N-terminus and one in the C-terminus, and a central Bub1-binding domain. During prophase and prometaphase, Bub3 localizes to the kinetochore before attachment to microtubules. In addition, taxol-induced formation of lagging chromosomes due to a delay of cell cycle progression increases the level of Bub3 co-localized with kinetochores, while correctly aligned chromosomes found in metaphase do not exhibit this co-localization. Thus, Bub3, in association with Bub1, may be important for sensing kinetochore attachment to microtubules during the prometaphase to metaphase transition.

Molecular Weight: 40 kDa

Pathways: Maintenance of Protein Location