Rabbit Recombinant Monoclonal PRAK/MK5 antibody. Suitable for WB, IP and reacts with Transfected cell lysate - Mouse, Human, Mouse, Rat samples.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
WB | IP | IHC-P | ICC/IF | Flow Cyt (Intra) | |
---|---|---|---|---|---|
Human | Tested | Tested | Not recommended | Not recommended | Not recommended |
Mouse | Tested | Tested | Not recommended | Not recommended | Not recommended |
Rat | Tested | Expected | Not recommended | Not recommended | Not recommended |
Transfected cell lysate - Mouse | Tested | Not recommended | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell lysate - Mouse | Dilution info 1/1000 | Notes - |
Species Human | Dilution info 1/1000 | Notes - |
Species Mouse | Dilution info 1/1000 | Notes - |
Species Rat | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/30 | Notes - |
Species Mouse | Dilution info 1/30 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell lysate - Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat, Transfected cell lysate - Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat, Transfected cell lysate - Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat, Transfected cell lysate - Mouse | Dilution info - | Notes - |
Tumor suppressor serine/threonine-protein kinase involved in mTORC1 signaling and post-transcriptional regulation. Phosphorylates FOXO3, ERK3/MAPK6, ERK4/MAPK4, HSP27/HSPB1, p53/TP53 and RHEB. Acts as a tumor suppressor by mediating Ras-induced senescence and phosphorylating p53/TP53. Involved in post-transcriptional regulation of MYC by mediating phosphorylation of FOXO3: phosphorylation of FOXO3 leads to promote nuclear localization of FOXO3, enabling expression of miR-34b and miR-34c, 2 post-transcriptional regulators of MYC that bind to the 3'UTR of MYC transcript and prevent MYC translation. Acts as a negative regulator of mTORC1 signaling by mediating phosphorylation and inhibition of RHEB. Part of the atypical MAPK signaling via its interaction with ERK3/MAPK6 or ERK4/MAPK4: the precise role of the complex formed with ERK3/MAPK6 or ERK4/MAPK4 is still unclear, but the complex follows a complex set of phosphorylation events: upon interaction with atypical MAPK (ERK3/MAPK6 or ERK4/MAPK4), ERK3/MAPK6 (or ERK4/MAPK4) is phosphorylated and then mediates phosphorylation and activation of MAPKAPK5, which in turn phosphorylates ERK3/MAPK6 (or ERK4/MAPK4). Mediates phosphorylation of HSP27/HSPB1 in response to PKA/PRKACA stimulation, inducing F-actin rearrangement.
MAP kinase-activated protein kinase 5, MAPK-activated protein kinase 5, MAPKAP kinase 5, MAPKAPK-5, Mapkapk5
Rabbit Recombinant Monoclonal PRAK/MK5 antibody. Suitable for WB, IP and reacts with Transfected cell lysate - Mouse, Human, Mouse, Rat samples.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
PRAK also known as MK5 is a serine/threonine protein kinase with a molecular weight of approximately 54 kDa. It is expressed in various tissues like the heart brain liver and skeletal muscle. PRAK belongs to the mitogen-activated protein kinase (MAPK) signaling pathway and gets activated by phosphorylation. Mechanically PRAK interacts with and phosphorylates other proteins which affects their activity and function playing an important role in regulating cellular processes such as stress response and the cell cycle.
PRAK/MK5 is a critical component in the regulation of cellular responses to stress stimuli. It is a participant in the activation loop of the MAPK p38 signaling pathway where it serves to influence the phosphorylation state and activity of downstream effectors. PRAK is not typically known to be part of a large complex but it interacts directly with other kinases to amplify or dampen the stress response signal. This allows cells to adapt to varied stress conditions by modulating gene expression apoptosis and other cell fate decisions.
PRAK/MK5 plays essential roles within the p38 MAPK and ERK signaling pathways. It closely associates with p38 MAPK which phosphorylates and activates PRAK enabling it to further disseminate the signal. In these pathways PRAK interplays with proteins such as MAPKAPK-2 and HSP27 facilitating diverse cellular activities like inflammation and differentiation. These signaling networks allow PRAK to serve as a link between external stimuli and internal cellular outcomes.
PRAK/MK5 influences cancer and cardiovascular diseases. Its activity connects closely with tumor suppression wherein it regulates actions involving proteins like p53 and HSP27 impacting cell cycle arrest and apoptosis. In cardiovascular contexts altered PRAK function can relate to heart disease since it regulates stress-induced cardiac responses. Understanding PRAK's interplay with these proteins sheds light on its potential as a therapeutic target for treating diseases related to stress signaling dysfunction.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Full details and terms and conditions can be found here:
Terms & Conditions.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 33442026).
This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
All lanes: Western blot - Anti-PRAK/MK5 antibody [EPR26844-237] (ab309533) at 1/1000 dilution
All lanes: Human brain tissue lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Developed using the ECL technique.
Observed band size: 50 kDa, 66 kDa
Exposure time: 180s
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 33442026).
In Western blot, ab309533 was shown to bind specifically to MAPKAPK5. Two bands were observed at 50 kDa and 66 kDa in wild-type L-929 cell lysates with whereas no signal observed at this size in MAPKAPK5 knockout cell line.
The lysates were kindly provided by an anonymous collaborator.
This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
All lanes: Western blot - Anti-PRAK/MK5 antibody [EPR26844-237] (ab309533) at 1/1000 dilution
Lane 1: Wild-type L-929 (mouse connective tissue fibroblast) whole cell lysate at 20 µg
Lane 2: MAPKAPK5 knockout L929 whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 50 kDa, 66 kDa
Exposure time: 125s
Blocking and diluting buffer and concentration: 5% NFDM/TBST
In Western blot, anti-His antibody (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution. Anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
Exposure time: 1 second
All lanes: Western blot - Anti-PRAK/MK5 antibody [EPR26844-237] (ab309533) at 1/1000 dilution
Lane 1: 293T cells transfected with an empty vector containi a His-tag whole cell lysate at 20 µg
Lane 2: 293T cells transfected with a Mouse MAPKAPK5 expression vector containi a His-tag whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 50 kDa
Exposure time: 1s
MAPKAPK5 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab309533 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab309533 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: Mouse brain tissue lysate
Lane 2: ab309533 IP in Mouse brain tissue lysate
Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab309533 in mouse brain tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 180 seconds
All lanes: Immunoprecipitation - Anti-PRAK/MK5 antibody [EPR26844-237] (ab309533) at 1/30 dilution
All lanes: Mouse brain tissue lysate
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 180s
MAPKAPK5 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab309533 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab309533 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2: ab309533 IP in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab309533 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 180 seconds
All lanes: Immunoprecipitation - Anti-PRAK/MK5 antibody [EPR26844-237] (ab309533) at 1/30 dilution
All lanes: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 180s
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 33442026).
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
Exposure time: 180 seconds
All lanes: Western blot - Anti-PRAK/MK5 antibody [EPR26844-237] (ab309533) at 1/1000 dilution
Lane 1: Mouse brain tissue lysate at 20 µg
Lane 2: Mouse heart tissue lysate at 20 µg
Lane 3: Mouse kidney tissue lysate at 20 µg
Lane 4: Rat brain tissue lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 50 kDa, 66 kDa
Exposure time: 180s
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 33442026).
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
Exposure time: 180 seconds
All lanes: Western blot - Anti-PRAK/MK5 antibody [EPR26844-237] (ab309533) at 1/1000 dilution
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 3: MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate at 20 µg
Lane 4: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 5: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 50 kDa, 66 kDa
Exposure time: 180s
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