Bub3 Recombinant Rabbit Monoclonal Antibody [JG38-81]
Catalog# ET7108-82
Bub3 Recombinant Rabbit Monoclonal Antibody [JG38-81]
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WB
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IF-Cell
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IHC-P
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FC
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IF-Tissue
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Human
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Mouse
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Rat
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Green monkey
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unconjugated
Overview
Product Name
Bub3 Recombinant Rabbit Monoclonal Antibody [JG38-81]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human Bub3 aa 2-50 / 328.
Species Reactivity
Human, Mouse, Rat, Green monkey
Validated Applications
WB, IF-Cell, IHC-P, FC, IF-Tissue
Molecular Weight
Predicted band size: 37 kDa
Positive Control
HeLa cell lysate, MCF7 cell lysate, COS-1 cell lysate, C2C12 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, PC-12 cell lysate, HeLa, C2C12, C6, human thyroid gland tissue, human colon cancer tissue, mouse fallopian tissue, rat testis tissue.
Conjugation
unconjugated
Clone Number
JG38-81
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000
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IF-Cell
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1:2,000
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IHC-P
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1:50-1:200
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FC
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1:1,000
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IF-Tissue
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1:50
Target
Function
BUB3 (budding uninhibited by benzimidazoles 3 homolog), also known as BUB3L or hBUB3, is a conserved component of the mitotic spindle assembly complex (MCC). It contains five WD repeat domains and forms cell cycle constitutive complexes with BUB1 and BUBR1. BUB3 is essential for the kinetochore localization of BUB1 and BUBR1. As a component of the MCC, BUB3 is involved in the essential spindle checkpoint pathway that operates during early embryogenesis. The spindle checkpoint pathway functions to postpone the initiation of anaphase until chromosomes are properly attached to the spindle. This acts to ensure accurate chromosome segregation. In addition, BUB3 plays a role in regulating the establishment of correct kinetochore-microtubule attachments. BUB3 is also thought to bind Tctex1L (or DYNLT3), a dynein light chain.
Background References
1. Seeley T W et al. Phosphorylation of human MAD1 by the BUB1 kinase in vitro. Biochem Biophys Res Commun 257:589-595 (1999).
2. Tang Z et al. Phosphorylation of Cdc20 by Bub1 provides a catalytic mechanism for APC/C inhibition by the spindle checkpoint. Mol Cell 16:387-397 (2004).
Sequence Similarity
Belongs to the WD repeat BUB3 family.
Post-translational Modification
Poly-ADP-ribosylated by PARP1.
Subcellular Location
Nucleus.
Synonyms
Aa2-050 antibody
AU019800 antibody
AU021329 antibody
AU043350 antibody
AW146323 antibody
BUB 3 antibody
BUB 3L antibody
BUB3 (budding uninhibited by benzimidazoles 3, yeast) homolog antibody
bub3 antibody
BUB3 budding uninhibited by benzimidazoles 3 homolog antibody
ExpandImages
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Western blot analysis of Bub3 on different lysates with Rabbit anti-Bub3 antibody (ET7108-82) at 1/2,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate
Lane 3: COS-1 cell lysate
Lane 4: C2C12 cell lysate
Lane 5: NIH/3T3 cell lysate
Lane 6: C6 cell lysate
Lane 7: PC-12 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 37 kDa
Observed band size: 40 kDa
Exposure time: 10 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7108-82) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of HeLa cells labeling Bub3 with Rabbit anti-Bub3 antibody (ET7108-82) at 1/2,000 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Bub3 antibody (ET7108-82) at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of C2C12 cells labeling Bub3 with Rabbit anti-Bub3 antibody (ET7108-82) at 1/2,000 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Bub3 antibody (ET7108-82) at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of C6 cells labeling Bub3 with Rabbit anti-Bub3 antibody (ET7108-82) at 1/2,000 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Bub3 antibody (ET7108-82) at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human thyroid gland tissue using anti-Bub3 antibody. Counter stained with hematoxylin.
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Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-Bub3 antibody. Counter stained with hematoxylin.
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Immunohistochemical analysis of paraffin-embedded mouse fallopian tissue using anti-Bub3 antibody. Counter stained with hematoxylin.
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Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-Bub3 antibody. Counter stained with hematoxylin.
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Flow cytometric analysis of HeLa cells labeling Bub3.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7108-82, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of C2C12 cells labeling Bub3.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7108-82, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"