Phospho-p38 (T180 + Y182) Rabbit Polyclonal Antibody
cat.: ER1903-01
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Tissue, FC
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at -20℃. Avoid repeated freeze/thaw cycles.
Storage buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: 42 kDa
Isotype: IgG
Immunogen: KLH conjugated Synthesised phosphopeptide derived from human p38 MAPK around the phosphorylation site of Thr180/Tyr182: M(p-T)G(p-Y)VA.
Positive control: Mouse muscle tissue, rat muscle tissue, rat brain tissue, mouse brain tissue, human placenta tissue, HepG2.
Subcellular location: Cytoplasm. Nucleus.
Recommended Dilutions:
  WB
  IHC-P
  IF-tissue
  FC
1:500-2000
1:100-500
1:100-500
1μg/Test
Uniprot #: SwissProt: Q16539 Human | P47811 Mouse | P70618 Rat
Alternative names: CSAID-binding protein Csaids binding protein CSBP CSBP1 CSBP2 CSPB1 Cytokine suppressive anti-inflammatory drug-binding protein EXIP MAP kinase 14 MAP kinase MXI2 MAP kinase p38 alpha MAPK 14 MAPK14 MAX-interacting protein 2 Mitogen-activated protein kinase 14 Mitogen-activated protein kinase p38 alpha MK14_HUMAN Mxi2 p38 p38 MAP kinase p38 mitogen activated protein kinase p38ALPHA p38alpha Exip PRKM14 PRKM15 RK SAPK2A Stress-activated protein kinase 2a
Images
ER1903-01_1.jpg Fig1: Sample:
Muscle (Mouse) Lysate at 40 ug
Muscle (Rat) Lysate at 40 ug
Primary: Anti-Phospho-P38 MAPK (Thr180 + Tyr182) (bs-0636R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kD
Observed band size: 42 kD
ER1903-01_2.jpg Fig2: Sample:
Kidney (Mouse) Lysate at 40 ug
Primary: Anti-Phospho-P38 MAPK (Thr180 + Tyr182) (bs-0636R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kD
Observed band size: 42 kD
ER1903-01_3.jpg Fig3: Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-P38 MAPK(Phospho-Thr180/Tyr182) Polyclonal Antibody, Unconjugated (bs-0636R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
ER1903-01_4.jpg Fig4: Tissue/cell: human placenta tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-P38 MAPK(Phospho-Thr180/Tyr182) Polyclonal Antibody, Unconjugated (bs-0636R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
ER1903-01_5.jpg Fig5: Tissue/cell: mouse brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-P38 MAPK(Phospho-Thr180/Tyr182) Polyclonal Antibody, Unconjugated (bs-0636R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
ER1903-01_6.jpg Fig6: Paraformaldehyde-fixed, paraffin embedded (rat brain tissue); 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°C for 30min; Antibody incubation with (P-P38 MAPK) Polyclonal Antibody, Unconjugated (bs-0636R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
ER1903-01_7.jpg Fig7: Blank control: HepG2(blue). Primary Antibody:Rabbit Anti-Phospho-P38 MAPK (Thr180 + Tyr182)antibody (bs-0636R,Green); Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions; Secondary Antibody: Goat anti-rabbit IgG-FITC(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.

Protocol
The cells were fixed with 2% paraformaldehyde for 10 min at 37℃. Primary antibody (bs-0636R, 1μg /1x10^6 cells) were incubated for 30 min at room temperature, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/FITC antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 40 min at room temperature. Acquisition of 20,000 events was performed.
ER1903-01_8.jpg Fig8: Blank control:MCF7.
Primary Antibody (green line): Rabbit Anti-Phospho-P38 MAPK (Thr180 + Tyr182) antibody (bs-0636R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1%PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.