BUB3 Recombinant Antibody

$IF(ICC) staining with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) 1:100 in A431 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.$

IF(ICC) staining with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) 1:100 in A431 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.

$IF(ICC) staining with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) 1:100 in A549 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.$

IF(ICC) staining with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) 1:100 in A549 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.

$IF(ICC) staining with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) 1:100 in LOVO cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.$

IF(ICC) staining with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) 1:100 in LOVO cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.

$Paraformaldehyde-fixed and paraffin-embedded Rat Testis tissue incubated with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) at 1:100, overnight at 4\u00b0C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.$

Paraformaldehyde-fixed and paraffin-embedded Rat Testis tissue incubated with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) at 1:100, overnight at 4\u00b0C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.

$Paraformaldehyde-fixed and paraffin-embedded Human Thyroid Gland tissue incubated with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) at 1:100, overnight at 4\u00b0C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.$

Paraformaldehyde-fixed and paraffin-embedded Human Thyroid Gland tissue incubated with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) at 1:100, overnight at 4\u00b0C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.

$Paraformaldehyde-fixed and paraffin-embedded Human Colon Cancer tissue incubated with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) at 1:100, overnight at 4\u00b0C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.$

Paraformaldehyde-fixed and paraffin-embedded Human Colon Cancer tissue incubated with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) at 1:100, overnight at 4\u00b0C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.

$Paraformaldehyde-fixed and paraffin-embedded Mouse Fallopian tissue incubated with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) at 1:100, overnight at 4\u00b0C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.$

Paraformaldehyde-fixed and paraffin-embedded Mouse Fallopian tissue incubated with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) at 1:100, overnight at 4\u00b0C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.

Flow cytometric analysis of PC-3M cells with BUB3 (24D1) Monoclonal Antibody (bsm-54413R) at a 1:100 dilution (purple) compared with an unlabeled control (cells without incubation with primary antibody; yellow).

$Paraformaldehyde-fixed, paraffin embedded Mouse testis; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse testis; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Mouse colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Mouse spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Rat spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Rat testis; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat testis; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Rat colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Human colon cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Human colon cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Human skin cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Human skin cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Human thyroid; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Human thyroid; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Human breast cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Human breast cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4\u00b0C, DAB staining.

$4% Paraformaldehyde-fixed HeLa (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Bub3) monoclonal Antibody, unconjugated (bsm-54413R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.$

4% Paraformaldehyde-fixed HeLa (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Bub3) monoclonal Antibody, unconjugated (bsm-54413R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.

$25 ug total protein per lane of various lysates (see on figure) probed with Bub3 monoclonal antibody, unconjugated (bsm-54413R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

25 ug total protein per lane of various lysates (see on figure) probed with Bub3 monoclonal antibody, unconjugated (bsm-54413R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

$The HeLa (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Cells stained with Primary Antibody for 30 min at room temperature,followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-Bub3 antibody (bsm-54413R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.$

The HeLa (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Cells stained with Primary Antibody for 30 min at room temperature,followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-Bub3 antibody (bsm-54413R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

$4% Paraformaldehyde-fixed HeLa (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Bub3) monoclonal Antibody, unconjugated (bsm-54413R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min . DAPI (blue, C02-04002) was used to stain the cell nuclei.$

4% Paraformaldehyde-fixed HeLa (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Bub3) monoclonal Antibody, unconjugated (bsm-54413R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min . DAPI (blue, C02-04002) was used to stain the cell nuclei.

Applications

WB
FCM
IHC-P
IF(IHC-P)
IF(ICC)
IP

Reactivity

Human
Mouse
Rat

Overview
Catalog #	bsm-54413r
Product Name	BUB3 Recombinant Antibody
Applications	WB FCM IHC-P IF(IHC-P) IF(ICC) IP
Reactivity	Human, Mouse, Rat
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Recombinant human Bub3
Clonality	Recombinant
Clone #	24D1
Isotype	IgG
Concentration	Lot Dependent
Purification	purified by Protein A.
Storage Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	9184
Swiss Prot	O43684
Subcellular location	Nucleus
Synonyms	BUB3L; hBUB3; CR201_G0012679; Aa2-050; BUB3_BOVIN; BUB3; BUB3_HUMAN; BUB3_MOUSE; WD repeat type I transmembrane protein A72.5; BUB3_PONAB; BUB3 mitotic checkpoint protein; BUB3 (budding uninhibited by benzimidazoles 3, yeast) homolog; budding uninhibited by benzimidazoles 3 homolog (yeast)
Background	Has a dual function in spindle-assembly checkpoint signaling and in promoting the establishment of correct kinetochore-microtubule (K-MT) attachments. Promotes the formation of stable end-on bipolar attachments. Necessary for kinetochore localization of BUB1. Regulates chromosome segregation during oocyte meiosis. The BUB1/BUB3 complex plays a role in the inhibition of anaphase-promoting complex or cyclosome (APC/C) when spindle-assembly checkpoint is activated and inhibits the ubiquitin ligase activity of APC/C by phosphorylating its activator CDC20. This complex can also phosphorylate MAD1L1.
Application Dilution
WB	1:300-5000
FCM	1:20-100
IHC-P	1:200-400
IF(IHC-P)	1:50-200
IF(ICC)	1:50-200
IP	1-2ug