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Aurora B Recombinant Rabbit Monoclonal Antibody [SC55-08]

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Specification

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Catalog# ET1610-25

Aurora B Recombinant Rabbit Monoclonal Antibody [SC55-08]

Application

  • WB

  • IHC-P

  • IP

  • IF-Cell

Reactivity

  • Human

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Aurora B Recombinant Rabbit Monoclonal Antibody [SC55-08]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Aurora B aa 1-50 / 344.

Species Reactivity

Human

Validated Applications

WB, IHC-P, IP, IF-Cell

Molecular Weight

Predicted band size: 39 kDa

Positive Control

Hela cell lysate, MCF-7 cell lysate, human tonsil tissue, HeLa.

Conjugation

unconjugated

Clone Number

SC55-08

RRID

AB_3069908

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

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:50-1:200

  • IF-Cell

  • 1:100

  • IP

  • 1-2μg/sample

Target

Function

Aurora related kinase-1 (ARK-1, STK15, Aurora2, Aik1) and -2 (ARK-2, STK12, Aurora1) are centrosome-associated serine/ threonine kinases that regulate centrosome separation, bipolar spindle assembly, and chromosome segregation during mitosis. ARK-1 and -2 are expressed in the nucleus and localize to distinct portions of mitotic machinery such as the centrosome, spindle poles (ARK-1), and midbody (ARK-2) during mitosis. ARK-1 and -2 transcripts are present at high levels in human thymus and fetal liver. ARK-1 protein has elevated expression in colon carcinoma lines (HT-29, SNU-C2B, COLO 205, SW480, 837 and 948) and accumulates during metaphase in HeLa cells. ARK-2 protein levels are maximal during both S and G2/M phases, whereas ARK-1 protein is degraded after G2/M via the ubiquitin-proteasome pathway. ARK-2 has a unique genetic loci relative to ARK-1, suggesting that these two kinases, with oncogenic potential, have different roles in cell cycle progression.

Background References

1. Petsalaki E & Zachos G Clks 1, 2 and 4 prevent chromatin breakage by regulating the Aurora B-dependent abscission checkpoint. Nat Commun 7:11451 (2016).

2. Jiang J et al. Cardiac myosin binding protein C regulates postnatal myocyte cytokinesis. Proc Natl Acad Sci U S A 112:9046-51 (2015).

Sequence Similarity

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. Aurora subfamily.

Tissue Specificity

High level expression seen in the thymus. It is also expressed in the spleen, lung, testis, colon, placenta and fetal liver. Expressed during S and G2/M phase and expression is up-regulated in cancer cells during M phase.

Post-translational Modification

The phosphorylation of Thr-232 requires the binding to INCENP and occurs by means of an autophosphorylation mechanism. Thr-232 phosphorylation is indispensable for the AURKB kinase activity.; Ubiquitinated by different BCR (BTB-CUL3-RBX1) E3 ubiquitin ligase complexes. Ubiquitinated by the BCR(KLHL9-KLHL13) E3 ubiquitin ligase complex, ubiquitination leads to removal from mitotic chromosomes and is required for cytokinesis. During anaphase, the BCR(KLHL21) E3 ubiquitin ligase complex recruits the CPC complex from chromosomes to the spindle midzone and mediates the ubiquitination of AURKB. Ubiquitination of AURKB by BCR(KLHL21) E3 ubiquitin ligase complex may not lead to its degradation by the proteasome.

Subcellular Location

Nucleus, Cytoplasm, Chromosome.

UNIPROT

SwissProt: Q96GD4 Human

SwissProt: O70126 Mouse

SwissProt: O55099 Rat

Synonyms

AIK2 antibody

AIM-1 antibody

AIM1 antibody

ARK-2 antibody

ARK2 antibody

AurB antibody

AURKB antibody

AURKB_HUMAN antibody

Aurora 1 antibody

Aurora and Ipl1 like midbody associated protein 1 antibody

Expand

Images

  • Western blot analysis of Aurora B on different lysates using anti-Aurora B antibody at 1/1,000 dilution.<br /><span style="font-weight: bold;">Positive control:</span> <br />Lane 1: Hela cell lysates <br />Lane 2: MCF-7 cell lysates

    Western blot analysis of Aurora B on different lysates using anti-Aurora B antibody at 1/1,000 dilution.
    Positive control:
    Lane 1: Hela cell lysates
    Lane 2: MCF-7 cell lysates

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />All lanes: Western blot analysis of Aurora B with anti-Aurora B antibody (<a href="/products/ET1610-25" style="font-weight: bold;text-decoration: underline;">ET1610-25</a>) at 1:500 dilution.<br />Lane 1: Wild-type Hela whole cell lysate (10 µg).<br />Lane 2: Aurora B knockdown Hela whole cell lysate (10 µg).<br /><br /><a href="/products/ET1610-25" style="font-weight: bold;text-decoration: underline;">ET1610-25</a> was shown to specifically react with Aurora B in wild-type Hela cells. Weakened band was observed when Aurora B knockdown sample was tested. Wild-type and Aurora B knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-25" style="font-weight: bold;text-decoration: underline;">ET1610-25</a>, 1:500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    All lanes: Western blot analysis of Aurora B with anti-Aurora B antibody (ET1610-25) at 1:500 dilution.
    Lane 1: Wild-type Hela whole cell lysate (10 µg).
    Lane 2: Aurora B knockdown Hela whole cell lysate (10 µg).

    ET1610-25 was shown to specifically react with Aurora B in wild-type Hela cells. Weakened band was observed when Aurora B knockdown sample was tested. Wild-type and Aurora B knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1610-25, 1:500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Aurora B antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Aurora B antibody. Counter stained with hematoxylin.

  • Immunocytochemistry analysis of HeLa cells labeling Aurora B with Rabbit anti-Aurora B antibody (<a href="/products/ET1610-25" style="font-weight: bold;text-decoration: underline;">ET1610-25</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Aurora B antibody (<a href="/products/ET1610-25" style="font-weight: bold;text-decoration: underline;">ET1610-25</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.

    Immunocytochemistry analysis of HeLa cells labeling Aurora B with Rabbit anti-Aurora B antibody (ET1610-25) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Aurora B antibody (ET1610-25) at 1/100 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.

  • Aurora B was immunoprecipitated from 0.2 mg HepG2 cell lysate with <a href="/products/ET1610-25" style="font-weight: bold;text-decoration: underline;">ET1610-25</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1610-25" style="font-weight: bold;text-decoration: underline;">ET1610-25</a> at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HepG2 cell lysate (input)<br />Lane 2: <a href="/products/ET1610-25" style="font-weight: bold;text-decoration: underline;">ET1610-25</a> IP in HepG2 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET1610-25" style="font-weight: bold;text-decoration: underline;">ET1610-25</a> in HepG2 cell lysate<br /><br />Blocking/Dilution buffer: primary antibody dilution (K1803)<br />Exposure time: 20 seconds; ECL: K1801

    Aurora B was immunoprecipitated from 0.2 mg HepG2 cell lysate with ET1610-25 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1610-25 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HepG2 cell lysate (input)
    Lane 2: ET1610-25 IP in HepG2 cell lysate
    Lane 3: Rabbit IgG instead of ET1610-25 in HepG2 cell lysate

    Blocking/Dilution buffer: primary antibody dilution (K1803)
    Exposure time: 20 seconds; ECL: K1801

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation