Aurora A Antibody - BSA Free

Novus Biologicals | Catalog # NBP1-51843

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Immunohistochemistry-Paraffin, Western Blot, Simple Western, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all Aurora A Primary Antibodies »

Product Specifications

Immunogen

A recombinant protein made to an N-terminal region of the human Aurora A protein (within residues 50-200). [Swiss-Prot O14965]

Localization

Centrosome, Spindle pole. Note: Detected at the neurite hillock in developing neurons. Localizes on centrosomes in interphase cells and at each spindle pole in mitosis.

Marker

Mitosis Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Aurora A Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: Aurora A Antibody - BSA Free [NBP1-51843]

Immunocytochemistry/ Immunofluorescence: Aurora A Antibody - BSA Free [NBP1-51843]

Immunocytochemistry/Immunofluorescence: Aurora A Antibody [NBP1-51843] - HeLa cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti- (NBP1-51843) at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution overnight at 4C and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Simple Western: Aurora A AntibodyBSA Free [NBP1-51843]

Simple Western: Aurora A AntibodyBSA Free [NBP1-51843]

Simple Western: Aurora A Antibody [NBP1-51843] - Simple Western lane view shows a specific band for Aurora A in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230kDa separation system.
Immunocytochemistry/ Immunofluorescence: Aurora A Antibody - BSA Free [NBP1-51843]

Immunocytochemistry/ Immunofluorescence: Aurora A Antibody - BSA Free [NBP1-51843]

Immunocytochemistry/Immunofluorescence: Aurora A Antibody [NBP1-51843] - IF Confocal analysis of HeLa cells using Aurora A antibody (NBP1-51843, 1:10). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
Western Blot: Aurora A AntibodyBSA Free [NBP1-51843]

Western Blot: Aurora A AntibodyBSA Free [NBP1-51843]

Western Blot: Aurora A Antibody [NBP1-51843] - WB detection of Aurora A in HeLa whole cell lysate.
Immunohistochemistry: Aurora A Antibody - BSA Free [NBP1-51843]

Immunohistochemistry: Aurora A Antibody - BSA Free [NBP1-51843]

Immunohistochemistry: Aurora A Antibody [NBP1-51843] - IHC staining of Aurora A in mouse brain.
Aurora A Antibody - BSA Free

Simple Western: Aurora A Antibody - BSA Free [NBP1-51843] -

Simple Western: Aurora A Antibody - BSA Free [NBP1-51843] - Analysis of protein expression post-drug treatment. (A) CHLA-10 and (B) TC-71 cells treated with drugs were assessed for changes in protein expression 24 h post-treatment via capillary electrophoresis-based Wes analysis. Increased protein levels of KIF11, p-KIF11Thr926 AURKA, and p-AURKAThr288 were observed for the drug combination group, whereas KIF15 levels were noticeably lower. Similarly, enhanced cleaved-PARP expression was observed with the combination treatment. The uncropped blots are shown in Figures S8 and S9. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/37894278 ), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Aurora A Antibody - BSA Free

Western Blot: Aurora A Antibody - BSA Free [NBP1-51843] -

An in silico bioinformatics screen identifies mitotic proteins essential for EWS growth. (A) The DepMap portal was used to access the RNA expression data across different cancer cell lines. Expression in Ewing sarcoma is highlighted in red. Capillary-based analysis of protein lysates from EWS cell lines indicating expression of (B) KIF11 and AURKA and (C) KIF15 and TPX2 protein levels. The uncropped blots are shown in Figure S3. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37894278), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Aurora A Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

2 ug/ml

Immunohistochemistry

1:200

Immunohistochemistry-Paraffin

1:200

Simple Western

1:200

Western Blot

1 - 2 ug/ml
Application Notes
This AURKA antibody is useful for Immunohistochemistry and Western blot, where a band is seen ~45 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in HeLa lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:200, apparent MW was 56 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 1 review rated 5 using NBP1-51843 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Aurora A

Aurora A is a cytoplasmic serine/threonine kinase that is critical in transitioning cells through mitosis and meiosis. Aurka acts through phosphorylating several spindle and tubulin proteins. Without Aurka, axon and neuron formation are negatively impacted. When these critical cellular processes are disrupted, there is a potential for tumorgenesis, and thus Aurka is implicated in many cancers, particularly those of the blood. In studies of angiofibromas, both MDM1 and Aurka were amplified. Overall, cell proliferation seems proportional to the expression of Aurka.

Alternate Names

AIK, ARK1, AURA, AURKA, BTAK, STK15, STK6, STK7

Entrez Gene IDs

6790 (Human); 20878 (Mouse)

Gene Symbol

AURKA

UniProt

O14965, P97477 (Mouse)

Additional Aurora A Products

Product Documents for Aurora A Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for Aurora A Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for Aurora A Antibody - BSA Free

Customer Reviews for Aurora A Antibody - BSA Free (1)

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  • Name: Bryan Tinsley
    Application: Immunocytochemistry
    Sample Tested:
    Species: Human
    Verified Customer | Posted 07/01/2014
    IF Confocal analysis of HeLa cells using Aurora A antibody (NBP1-51843, 1:10).
    Aurora A Antibody - BSA Free NBP1-51843

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Protocols

View specific protocols for Aurora A Antibody - BSA Free (NBP1-51843):

Aurora A Antibody:
Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.

Aurora A Antibody:
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

*Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

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FAQs for Aurora A Antibody - BSA Free

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  • Q: Aurora A Antibody NBP1-51843 says it is compatible for IF at 1:10-1:2000. Do you have a more accurate concentration for paraffin embedded tissue? 5um sections. Thanks.

    A: IF/ICC is our abbreviation for immunofluorescencestaining of cells. For IHC with paraffin-embedded tissue the recommended starting dilution is 1:50. Antigen retrieval with sodium citrate buffer (pH 6.0) is recommended for this antibody when performing IHC-P. Please accept my apologies for any confusion, and feel free to contact me should you have any further questions.

  • Q: What are the better positive controls for Aurora A Antibody (NBP1-51843) in the immunohistochemistry reaction, dilution used and antigen retrieval?

    A: The dilution we recommend for IHC is 1:50 and here is our protocol for Antigen Unmasking: Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes. As far as positive control is concerned, Aurora A is highly expressed in testis, colon, ovarian, prostate, neuroblastoma, breast and cervical cancer, so that you may include one of these tissues as your positive controls.

  • Q: Aurora A Antibody NBP1-51843 says it is compatible for IF at 1:10-1:2000. Do you have a more accurate concentration for paraffin embedded tissue? 5um sections. Thanks.

    A: IF/ICC is our abbreviation for immunofluorescencestaining of cells. For IHC with paraffin-embedded tissue the recommended starting dilution is 1:50. Antigen retrieval with sodium citrate buffer (pH 6.0) is recommended for this antibody when performing IHC-P. Please accept my apologies for any confusion, and feel free to contact me should you have any further questions.

  • Q: What are the better positive controls for Aurora A Antibody (NBP1-51843) in the immunohistochemistry reaction, dilution used and antigen retrieval?

    A: The dilution we recommend for IHC is 1:50 and here is our protocol for Antigen Unmasking: Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes. As far as positive control is concerned, Aurora A is highly expressed in testis, colon, ovarian, prostate, neuroblastoma, breast and cervical cancer, so that you may include one of these tissues as your positive controls.

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