bsm-54413R [Primary Antibody]
BUB3 Recombinant Antibody
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Host: Rabbit

Target Protein: Bub3

Clonality: Recombinant

Isotype: IgG

Entrez Gene: 9184

Swiss Prot: O43684

Source: Recombinant protein corresponding to the N-terminus of human Bub3.

Purification: Purified by Protein A.

Storage Buffer: 0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol

Storage: Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.

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.

Size: 100µL

Concentration: 1ug/ul

Predicted Molecular Weight: 37


Cross Reactive Species: Human
Mouse
Rat

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

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 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°C, 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°C, 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°C, 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°C, 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°C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4°C, 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°C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4°C, 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°C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4°C, 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°C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4°C, 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°C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4°C, 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°C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4°C, 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°C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4°C, 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°C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4°C, 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°C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4°C, 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°C for 30min; Antibody incubation with BUB3 (24D1) Monoclonal Antibody, Unconjugated (bsm-54413R) at 1:200 overnight at 4°C, 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°C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37°C 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°C 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℃,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.