PRODUCT CODE: ET1609-22

Aurora A Recombinant Rabbit Monoclonal Antibody [ST46-07] (ET1609-22)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • FC

REACTIVITY

  • Human

  • Mouse

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Western blot analysis of Aurora A on K562 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1609-22, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of Aurora A on K562 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1609-22, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.
  • ICC staining of Aurora A in SKOV-3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-22, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Aurora A in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-22, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Aurora A in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-22, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-Aurora A antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-22, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of Aurora A was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1609-22, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of Aurora A on K562 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1609-22, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IF

  • FC

REACTIVITY

  • Human

  • Mouse

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Aurora A Recombinant Rabbit Monoclonal Antibody [ST46-07] (ET1609-22)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

SKOV-3, NIH/3T3, Hela.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

ST46-07

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

46 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • FC

  • 1:50-1:100

  • ICC/IF

  • 1:100-1:500

TARGET

UNIPROT #

PROTEIN NAME

Aurora A

SYNONYMS

AIK antibody; ARK-1 antibody; ARK1 antibody; AURA antibody; Aurka antibody; Aurora 2 antibody; Aurora A antibody; Aurora kinase A antibody; Aurora-related kinase 1 antibody; Aurora/IPL1 like kinase antibody; Aurora/IPL1-related kinase 1 antibody; AURORA2 antibody; Breast tumor-amplified kinase antibody; BTAK antibody; hARK1 antibody; IAK antibody; IPL1 related kinase antibody; MGC34538 antibody; OTTHUMP00000031340 antibody; OTTHUMP00000031341 antibody; OTTHUMP00000031342 antibody; OTTHUMP00000031343 antibody; OTTHUMP00000031344 antibody; OTTHUMP00000031345 antibody; OTTHUMP00000166071 antibody; OTTHUMP00000166072 antibody; PPP1R47 antibody; Protein phosphatase 1, regulatory subunit 47 antibody; Serine/threonine kinase 15 antibody; Serine/threonine kinase 6 antibody; Serine/threonine-protein kinase 15 antibody; Serine/threonine-protein kinase 6 antibody; Serine/threonine-protein kinase aurora-A antibody; STK15 antibody; STK6 antibody; STK6_HUMAN antibody; STK7 antibody

SEQUENCE SIMILARITIES

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

TISSUE SPECIFICITY

Highly expressed in testis and weakly in skeletal muscle, thymus and spleen. Also highly expressed in colon, ovarian, prostate, neuroblastoma, breast and cervical cancer cell lines.

POST-TRANSLATIONAL MODIFICATION

Activated by phosphorylation at Thr-288; this brings about a change in the conformation of the activation segment. Phosphorylation at Thr-288 varies during the cell cycle and is highest during M phase. Autophosphorylated at Thr-288 upon TPX2 binding. Thr-288 can be phosphorylated by several kinases, including PAK and PKA. Protein phosphatase type 1 (PP1) binds AURKA and inhibits its activity by dephosphorylating Thr-288 during mitosis. Phosphorylation at Ser-342 decreases the kinase activity. PPP2CA controls degradation by dephosphorylating Ser-51 at the end of mitosis.; Ubiquitinated by the E3 ubiquitin-protein ligase complex SCF(FBXL7) during mitosis, leading to its degradation by the proteasome (By similarity). Ubiquitinated by CHFR, leading to its degradation by the proteasome (By similarity). Ubiquitinated by the anaphase-promoting complex (APC), leading to its degradation by the proteasome. Ubiquitinated by the CUL3-KLHL18 ligase leading to its activation at the centrosome which is required for initiating mitotic entry. Ubiquitination mediated by CUL3-KLHL18 ligase does not lead to its degradation by the proteasome.

SUBCELLULAR LOCATION

Cytoplasm, Cytoskeleton, Centrosome, Nucleus.

FUNCTION

Activation of the oncogenic protein kinase Aurora A regulates meiotic and mitotic cell cycles in eukaryotic cells. Specifically, Aurora A plays a key role in G2/M progression. Activation occurs via autophosphorylation, and while 14 sites are subject to this, only the threonine residue at position 295 is required for activity. Though autophosphorylation mediates activation, a number of other proteins influence activation, including the spindle assembly factor TPX2 and p53

CITATIONS

  • Li, Ran et al.

    Involvement of polo-like kinase 1 (Plk1) in mitotic arrest by inhibition of mitogen-activated protein kinase-extracellular signal-regulated kinase-ribosomal S6 kinase 1 (MEK-ERK-RSK1) cascade. | The Journal of Biological Chemistry [2012]