p38 alpha / MAPK14 Recombinant Rabbit Monoclonal Antibody [JF55-07]

Specification

50μl

100μl

Quota

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Contact: Sanry CHEN

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Phone: 852-35650223

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Catalog# ET1702-65

p38 alpha / MAPK14 Recombinant Rabbit Monoclonal Antibody [JF55-07]

Application

WB
IF-Cell
IF-Tissue
IHC-P
FC

Reactivity

Human
Mouse
Rat

Conjugation

unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

p38 alpha / MAPK14 Recombinant Rabbit Monoclonal Antibody [JF55-07]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within C-terminal human MAPK14.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, FC

Molecular Weight

Predicted band size: 41 kDa

Positive Control

HeLa cell lysate, Jurkat cell lysate, HEK-293 cell lysate, NIH/3T3 cell lysate, Neuro-2a cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, C6 cell lysate, THP-1 cell lysate, C2C12 cell lysate, Rat kidney tissue lysate, Mouse kidney tissue lysate, RAW264.7, PC-12, HeLa, mouse heart tissue.

Conjugation

unconjugated

Clone Number

JF55-07

RRID

AB_3073466

Product Features

Form

Liquid

Concentration

1 mg/mL.

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

WB
1:2,000-1:5,000

IF-Cell
1:100

IF-Tissue
1:50-1:200

IHC-P
1:50-1:200

FC
1:1,000

Applications in Publications

WB	See 48 publications below

Species in Publications

Mouse	See 24 publications below
Human	See 15 publications below
Rat	See 4 publications below
Pig	See 3 publications below
Chicken	See 1 publications below
Sheep	See 1 publications below
M. Oryzae	See 1 publications below

Target

Function

MAP (mitogen-activated protein) kinases play a significant role in many biological processes, including cell adhesion and spreading, cell differentiation and apoptosis. p38α, p38β and p38γ, also known as MAPK14, MAPK11 and MAPK12, respectively, each contain one protein kinase domain and belong to the MAP kinase family. Expressed in different areas throughout the body with common expression patterns in heart, p38 proteins use magnesium as a cofactor to catalyze the ATP-dependent phosphorylation of target proteins. Via their catalytic activity, p38α, p38β and p38γ are involved in a variety of events throughout the cell, including signal transduction pathways, cytokine production and cell proliferation and differentiation. The p38 proteins are subject to phosphoryation on Thr and Tyr residues, an event which is thought to activate the phosphorylated protein.

Background References

1. Xiao Y et al. Effect of a-linolenic acid-modified low molecular weight chondroitin sulfate on atherosclerosis in apoE-deficient mice. Biochim Biophys Acta 1860:2589-97 (2016).

2. Lv L et al. WT1-AS promotes cell apoptosis in hepatocellular carcinoma through down-regulating of WT1. J Exp Clin Cancer Res 34:119 (2015).

Sequence Similarity

Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.

Tissue Specificity

Brain, heart, placenta, pancreas and skeletal muscle. Expressed to a lesser extent in lung, liver and kidney.

Post-translational Modification

Dually phosphorylated on Thr-180 and Tyr-182 by the MAP2Ks MAP2K3/MKK3, MAP2K4/MKK4 and MAP2K6/MKK6 in response to inflammatory citokines, environmental stress or growth factors, which activates the enzyme. Dual phosphorylation can also be mediated by TAB1-mediated autophosphorylation. TCR engagement in T-cells also leads to Tyr-323 phosphorylation by ZAP70. Dephosphorylated and inactivated by DUPS1, DUSP10 and DUSP16. PPM1D also mediates dephosphorylation and inactivation of MAPK14.; Acetylated at Lys-53 and Lys-152 by KAT2B and EP300. Acetylation at Lys-53 increases the affinity for ATP and enhances kinase activity. Lys-53 and Lys-152 are deacetylated by HDAC3.; Ubiquitinated. Ubiquitination leads to degradation by the proteasome pathway.

Subcellular Location

Nucleus, Cytoplasm.

UNIPROT

SwissProt: Q16539 Human

SwissProt: P47811 Mouse

SwissProt: P70618 Rat

Synonyms

CSAID-binding protein antibody

Csaids binding protein antibody

CSBP antibody

CSBP1 antibody

CSBP2 antibody

CSPB1 antibody

Cytokine suppressive anti-inflammatory drug-binding protein antibody

EXIP antibody

MAP kinase 14 antibody

MAP kinase MXI2 antibody

Expand

Images

Western blot analysis of p38 alpha / MAPK14 on different lysates with Rabbit anti-p38 alpha / MAPK14 antibody (ET1702-65) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: Jurkat cell lysate
Lane 3: HEK-293 cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: Neuro-2a cell lysate
Lane 6: RAW264.7 cell lysate
Lane 7: PC-12 cell lysate
Lane 8: C6 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 41 kDa
Observed band size: 38 kDa

Exposure time: 3 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-65) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Knockout (KO)

All lanes: Western blot analysis of p38 alpha / MAPK14 with anti-p38 alpha / MAPK14 antibody[JF55-07] (ET1702-65) at 1:500 dilution.
Lane 1: Wild-type Hela whole cell lysate (10 µg).
Lane 2: p38 alpha / MAPK14 knockout Hela whole cell lysate (10 µg).

ET1702-65 was shown to specifically react with p38 alpha / MAPK14 in wild-type Hela cells. No band was observed when p38 alpha / MAPK14 knockout sample was tested. Wild-type and p38 alpha / MAPK14 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1702-65, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
Western blot analysis of p38 alpha / MAPK14 on different lysates with Rabbit anti-p38 alpha / MAPK14 antibody (ET1702-65) at 1/1,000 dilution.

Lane 1: THP-1 cell lysate (10 µg/Lane)
Lane 2: C2C12 cell lysate (10 µg/Lane)
Lane 3: PC-12 cell lysate (10 µg/Lane)
Lane 4: Rat kidney tissue lysate (20 µg/Lane)
Lane 5: Mouse kidney tissue lysate (20 µg/Lane)

Predicted band size: 41 kDa
Observed band size: 38 kDa

Exposure time: 3 minutes 10 seconds;

10% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-65) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of RAW264.7 cells labeling p38 alpha / MAPK14 with Rabbit anti-p38 alpha / MAPK14 antibody (ET1702-65) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-p38 alpha / MAPK14 antibody (ET1702-65) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of PC-12 cells labeling p38 alpha / MAPK14 with Rabbit anti-p38 alpha / MAPK14 antibody (ET1702-65) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-p38 alpha / MAPK14 antibody (ET1702-65) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Flow cytometric analysis of RAW264.7 cells labeling p38 alpha / MAPK14.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-65, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Flow cytometric analysis of HeLa cells labeling p38 alpha / MAPK14.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-65, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-p38 alpha / MAPK14 antibody (ET1702-65) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1702-65) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.