PA5-17689

antibody from Invitrogen Antibodies

Targeting: MAPK7 BMK1, ERK5, PRKM7

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-17689

Provider

Invitrogen Antibodies

Product name

ERK5 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody.

Concentration

23 µg/mL

References

Submitted references

Identification of Novel RAS Signaling Therapeutic Vulnerabilities in Diffuse Intrinsic Pontine Gliomas.

Koncar RF, Dey BR, Stanton AJ, Agrawal N, Wassell ML, McCarl LH, Locke AL, Sanders L, Morozova-Vaske O, Myers MI, Hamilton RL, Carcaboso AM, Kohanbash G, Hu B, Amankulor NM, Felker J, Kambhampati M, Nazarian J, Becher OJ, James CD, Hashizume R, Broniscer A, Pollack IF, Agnihotri S
Cancer research 2019 Aug 15;79(16):4026-4041

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Erk5 was performed by loading 30 µg of HEK293T whole cell lysates per well onto an SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat dry milk in TBST for 1 hour at room temperature. The membrane was probed with an Erk5 polyclonal antibody (Product # PA5-17689) at a dilution of 1:500 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated goat anti-rabbit IgG at a dilution of 1:40,000 for 1 hour at room temperature. Detection was performed using ECL substrate. Data courtesy of the Innovators Program.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of Mitogen-activated protein kinase 7 was achieved by transfecting DU 145 with Mitogen-activated protein kinase 7 specific siRNAs (Silencer® select Product # s11149, s11151). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Mitogen-activated protein kinase 7 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with ERK5 Polyclonal Antibody (Product # PA5-17689, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:6000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Mitogen-activated protein kinase 7.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-ERK5 Polyclonal Antibody (Product # PA5-17689) and a 115 kDa band corresponding to Mitogen-activated protein kinase 7 was observed across cell lines and tissue tested. Upon EGF and sorbitol treatment in HeLa cells, there was a shift in the band size which corresponds to phosphorylated form of ERK5, since it is an autophosphorylated protein. Whole cell extracts (30 µg lysate) of (Fig.a) HeLa (Lane 1), HeLa serum starved for overnight and treated with EGF (100 ng/mL for 15 min) (Lane 2), HeLa treated with Sorbitol (0.5M for 30 min) (Lane 3) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX); (Fig.b) PC-3 (Lane 1), DU 145 (Lane 2), NIH:OVCAR-3 (Lane 3), SK-O-V3 (Lane 4), NIH/3T3 (Lane 5), Mouse Testis (Lane 6), Mouse Ovary (Lane 7). 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-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:6000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).