ERK5 (8D8) Monoclonal Antibody

HeLa cell lysates probed with ERK5 (8D8) Monoclonal Antibody (bsm-52069R) at 1:1000 overnight at 4˚C. Followed by a conjugated secondary antibody.

MCF-7 cells were stained with ERK5 (8D8) Monoclonal Antibody (bsm-52069R) at [1:200] incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.

$MCF7 cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with ERK5 (8D8) Monoclonal Antibody(bsm-52069R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

MCF7 cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with ERK5 (8D8) Monoclonal Antibody(bsm-52069R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

$Paraformaldehyde-fixed and paraffin-embedded Rat testis tissue incubated with ERK5 (8D8) Monoclonal Antibody (bsm-52069R) at 1:200, overnight at 4\u00b0C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.$

Paraformaldehyde-fixed and paraffin-embedded Rat testis tissue incubated with ERK5 (8D8) Monoclonal Antibody (bsm-52069R) at 1:200, overnight at 4\u00b0C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.

$Paraformaldehyde-fixed, paraffin embedded Rat testis; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with ERK5 (8D8) Monoclonal Antibody, Unconjugated (bsm-52069R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat testis; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with ERK5 (8D8) Monoclonal Antibody, Unconjugated (bsm-52069R) at 1:200 overnight at 4\u00b0C, DAB staining.

Lane 1: Mouse Testis lysates; Lane 2: Human Hela cell lysates probed with ERK5 monoclonal Antibody, Unconjugated (bsm-52069R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

$Blocking buffer: 5% NFDM\/TBST\r\nPrimary Ab dilution: 1:2000\r\nPrimary Ab incubation condition: 2 hours at \r\nroom temperature\r\nSecondary Ab: Goat Anti-Rabbit IgG H&L \r\n(HRP)\r\nLysate: 1: MCF-7, 2: 293T, 3: NIH\/3T3, 4: PC-\r\n12\r\nProtein loading quantity: 20 \u03bcg\r\nExposure time: 60 s\r\nPredicted MW: 88 kDa\r\nObserved MW: 115 kDa$

Blocking buffer: 5% NFDM\/TBST\r\nPrimary Ab dilution: 1:2000\r\nPrimary Ab incubation condition: 2 hours at \r\nroom temperature\r\nSecondary Ab: Goat Anti-Rabbit IgG H&L \r\n(HRP)\r\nLysate: 1: MCF-7, 2: 293T, 3: NIH\/3T3, 4: PC-\r\n12\r\nProtein loading quantity: 20 \u03bcg\r\nExposure time: 60 s\r\nPredicted MW: 88 kDa\r\nObserved MW: 115 kDa

Applications

WB
FCM
IHC-P
IF(ICC)

Reactivity

Human
Mouse
Rat

Overview
Catalog #	bsm-52069R
Product Name	ERK5 (8D8) Monoclonal Antibody
Applications	WB, FCM, IHC-P, IF(ICC)
Reactivity	Human, Mouse, Rat
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Recombinant human ERK5 protein, around C-terminal 200aa.
Clonality	Monoclonal
Clone #	8D8
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	Aqueous buffered solution containing 1xTBS (pH7.4), 1%BSA, 40%Glycerol and 0.05% Sodium Azide.
Storage Condition	Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	5598
Swiss Prot	Q13164
Synonyms	BMK1; ERK4; ERK5; PRKM7; Mitogen-activated protein kinase 7; MAP kinase 7; MAPK 7; Big MAP kinase 1; BMK-1; Extracellular signal-regulated kinase 5; ERK-5; MAPK7
Background	Plays a role in various cellular processes such as proliferation, differentiation and cell survival. The upstream activator of MAPK7 is the MAPK kinase MAP2K5. Upon activation, it translocates to the nucleus and phosphorylates various downstream targets including MEF2C. EGF activates MAPK7 through a Ras-independent and MAP2K5-dependent pathway. May have a role in muscle cell differentiation. May be important for endothelial function and maintenance of blood vessel integrity. MAP2K5 and MAPK7 interact specifically with one another and not with MEK1/ERK1 or MEK2/ERK2 pathways. Phosphorylates SGK1 at Ser-78 and this is required for growth factor-induced cell cycle progression. Involved in the regulation of p53/TP53 by disrupting the PML-MDM2 interaction.
Application Dilution
WB	1:300-5000
FCM	1:20-100
IHC-P	1:200-400
IF(ICC)	1:50-200