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p38 alpha / MAPK14 Recombinant Rabbit Monoclonal Antibody [SR43-04]

Usd: 385

Specification

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Catalog# ET1602-26

p38 alpha / MAPK14 Recombinant Rabbit Monoclonal Antibody [SR43-04]

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

p38 alpha / MAPK14 Recombinant Rabbit Monoclonal Antibody [SR43-04]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human p38 aa 160-200.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 41 kDa

Positive Control

Hela cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, THP-1 cell lysate, Jurkat cell lysate, C2C12 cell lysate, rat kidney tissue lysate, mouse kidney tissue lysate, HeLa, RAW264.7, C6.

Conjugation

unconjugated

Clone Number

SR43-04

RRID

AB_3069639

Product Features

Form

Liquid

Concentration

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

  • 1:1,000

  • IF-Cell

  • 1:100-1:500

  • FC

  • 1:1,000

Target

Function

p38 mitogen-activated protein kinases are a class of mitogen-activated protein kinases (MAPKs) that are responsive to stress stimuli, such as cytokines, ultraviolet irradiation, heat shock, and osmotic shock, and are involved in cell differentiation, apoptosis and autophagy. Persistent activation of the p38 MAPK pathway in muscle satellite cells (muscle stem cells) due to ageing, impairs muscle regeneration. p38 MAP Kinase (MAPK), also called RK or CSBP (Cytokinin Specific Binding Protein), is the mammalian orthologue of the yeast Hog1p MAP kinase, which participates in a signaling cascade controlling cellular responses to cytokines and stress. Four p38 MAP kinases, p38-α (MAPK14), -β (MAPK11), -γ (MAPK12 / ERK6), and -δ (MAPK13 / SAPK4), have been identified. Similar to the SAPK/JNK pathway, p38 MAP kinase is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharides (LPS), ultraviolet light, and growth factors. MKK3 and SEK activate p38 MAP kinase by phosphorylation at Thr-180 and Tyr-182.

Background References

1. Yan T et al. Luteolin inhibits behavioral sensitization by blocking methamphetamine-induced MAPK pathway activation in the caudate putamen in mice. PLoS One 9:e98981 (2014).

2. Pagliara V et al. Protease Nexin-1 affects the migration and invasion of C6 glioma cells through the regulation of urokinase Plasminogen Activator and Matrix Metalloproteinase-9/2. Biochim Biophys Acta 1843:2631-44 (2014).

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

Cytoplasm, Nucleus.

UNIPROT

SwissProt: Q16539 Human

SwissProt: P47811 Mouse

SwissProt: P70618 Rat

Synonyms

CSAID Binding Protein 1 antibody

CSAID binding protein antibody

CSAID-binding protein antibody

Csaids binding protein antibody

CSBP 1 antibody

CSBP 2 antibody

CSBP antibody

CSBP1 antibody

CSBP2 antibody

CSPB1 antibody

Expand

Images

  • Western blot analysis of p38 alpha / MAPK14 on different lysates with Rabbit anti-p38 alpha / MAPK14 antibody (<a href="/products/ET1602-26" style="font-weight: bold;text-decoration: underline;">ET1602-26</a>) at 1/1,000 dilution.<br /><br />Lane 1: Hela cell lysate<br />Lane 2: NIH/3T3 cell lysate<br />Lane 3: PC-12 cell lysate<br />Lane 4: THP-1 cell lysate<br />Lane 5: Jurkat cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 41 kDa<br />Observed band size: 38 kDa<br /><br />Exposure time: 2 minutes;<br />12% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1602-26" style="font-weight: bold;text-decoration: underline;">ET1602-26</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 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 (ET1602-26) at 1/1,000 dilution.

    Lane 1: Hela cell lysate
    Lane 2: NIH/3T3 cell lysate
    Lane 3: PC-12 cell lysate
    Lane 4: THP-1 cell lysate
    Lane 5: Jurkat cell lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 2 minutes;
    12% 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 (ET1602-26) 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 1:300,000 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 (<a href="/products/ET1602-26" style="font-weight: bold;text-decoration: underline;">ET1602-26</a>) at 1/1,000 dilution.<br /><br />Lane 1: THP-1 cell lysate (10 µg/Lane)<br />Lane 2: C2C12 cell lysate (10 µg/Lane)<br />Lane 3: PC-12 cell lysate (10 µg/Lane)<br />Lane 4: Rat kidney tissue lysate (20 µg/Lane)<br />Lane 5: Mouse kidney tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 41 kDa<br />Observed band size: 38 kDa<br /><br />Exposure time: 3 minutes 10 seconds;<br />10% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1602-26" style="font-weight: bold;text-decoration: underline;">ET1602-26</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) 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 (ET1602-26) 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 (ET1602-26) 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of p38 alpha / MAPK14 on different lysates with Rabbit anti-p38 alpha / MAPK14 antibody (<a href="/products/ET1602-26" style="font-weight: bold;text-decoration: underline;">ET1602-26</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549-WT cell lysate<br />Lane 2: A549-KD p38 alpha / MAPK14 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 41 kDa<br />Observed band size: 38 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1602-26" style="font-weight: bold;text-decoration: underline;">ET1602-26</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of p38 alpha / MAPK14 on different lysates with Rabbit anti-p38 alpha / MAPK14 antibody (ET1602-26) at 1/2,000 dilution.

    Lane 1: A549-WT cell lysate
    Lane 2: A549-KD p38 alpha / MAPK14 cell lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 3 minutes; ECL: K1801;

    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 (ET1602-26) at 1/2,000 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of HeLa cells treated with UV for 30 minutes then recover 30 minutes labeling p38 alpha / MAPK14 with Rabbit anti-p38 alpha / MAPK14 antibody (<a href="/products/ET1602-26" style="font-weight: bold;text-decoration: underline;">ET1602-26</a>) at 1/500 dilution.<br /><br />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 (<a href="/products/ET1602-26" style="font-weight: bold;text-decoration: underline;">ET1602-26</a>) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of HeLa cells treated with UV for 30 minutes then recover 30 minutes labeling p38 alpha / MAPK14 with Rabbit anti-p38 alpha / MAPK14 antibody (ET1602-26) at 1/500 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 (ET1602-26) at 1/500 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 RAW264.7 cells labeling p38 alpha / MAPK14 with Rabbit anti-p38 alpha / MAPK14 antibody (<a href="/products/ET1602-26" style="font-weight: bold;text-decoration: underline;">ET1602-26</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/ET1602-26" style="font-weight: bold;text-decoration: underline;">ET1602-26</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of RAW264.7 cells labeling p38 alpha / MAPK14 with Rabbit anti-p38 alpha / MAPK14 antibody (ET1602-26) 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 (ET1602-26) 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 C6 cells labeling p38 alpha / MAPK14 with Rabbit anti-p38 alpha / MAPK14 antibody (<a href="/products/ET1602-26" style="font-weight: bold;text-decoration: underline;">ET1602-26</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/ET1602-26" style="font-weight: bold;text-decoration: underline;">ET1602-26</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of C6 cells labeling p38 alpha / MAPK14 with Rabbit anti-p38 alpha / MAPK14 antibody (ET1602-26) 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 (ET1602-26) 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 HeLa cells labeling p38 alpha / MAPK14.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1602-26" style="font-weight: bold;text-decoration: underline;">ET1602-26</a>, 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&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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 (ET1602-26, 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).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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