MA5-15321

antibody from Invitrogen Antibodies

Targeting: AURKB Aik2, AIM-1, ARK2, AurB, IPL1, PPP1R48, STK12, STK5

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-15321

Provider

Invitrogen Antibodies

Product name

Aurora B Monoclonal Antibody (13E8D3)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

13E8D3

Vial size

100 µL

Concentration

Conc. Not Determined

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Klotho Exerts an Emerging Role in Cytokinesis.

Sun CY, Chou CY, Hsieh YY, Lo KC, Liou YL, Chen YH
Genes 2020 Sep 4;11(9)

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of AURKB using a AURKB monoclonal antibody (Product # MA5-15321) against a truncated AURKB recombinant protein (1) and SKN-SH cell lysate (2).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Aurora B Monoclonal Antibody (13E8D3) (Product # MA5-15321) and a 39 kDa band corresponding to Aurora B was observed across cell lines tested and increased upon Nocodazole treatment in HeLa. Whole cell extracts (30 µg lysate) of NTERA-2 (Lane 1), A-549 (Lane 2), T-47D (Lane 3), HEK (Lane 4), HeLa (Lane 5), HeLa treated with Nocodazole (200ng/ml for 24 hours (Lane 6) and THP-1 (Lane 7) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)..

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Aurora B was performed using 70% confluent log phase HeLa and T-47D cells. The cells were fixed with 4% Paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 10 minutes at room temperature. The cells were labeled with Aurora B Monoclonal Antibody (13E8D3) (Product # MA5-15321) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175, 1:2000 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing plasma membrane localization. Panel e represents T-47D cells having low expression of Aurora B. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 6 Klotho depletion impairs Aurora kinase activation. ( A - C ) Immunoblot analysis of Klotho, and Aurora kinase A and B with the protein extracts of transfected HeLa cells, respectively. ( D , E ) Immunoblot analysis of Klotho and Aurora kinase B with the protein extracts of HeLa cells treated with nocodazole for 16 h, respectively. Each treatment for ( B , C ) was repeated in triplicate, and the statistic results are plotted (means +- SEM; one-way ANOVA; * p < 0.05; ** p < 0.01; KL--Klotho; Aura A--Aurora kinase A; Aura B--Aurora kinase B).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 7 3D reconstruction of the cytokinesis bridge. Intrinsic Klotho, Aurora kinase B, and citron kinase of cultured HeLa cells analyzed by immunofluorescence. ( A ) Results of immunofluorescence staining for Klotho and Aurora kinase B. ( B ) 3D structure of Klotho and Aurora kinase B in the cytokinesis bridge. ( C ) 3D structure of Klotho and citron kinase in the cytokinesis bridge. ( D ) Midbody architecture related with Klotho, Aurora kinase B, and citron kinase (microscopy: x400).