Anti-p38 alpha/MAPK14 Antibody

• Catalog Number: orb1098071
• Category: Antibodies
• Description: Anti-p38 alpha/MAPK14 Antibody. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IHC, IP, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human p38 alpha/MAPK14, identical to the related mouse and rat sequences.
• Antibody Type: Primary Antibody
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 38 kDa
• UniProt ID: Q16539
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: Mannose-6-phosphate isomerase; 5.3.1.8; Phosphohex
• Note: For research use only
• Application notes: Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat Immunohistochemistry, 2-5 μg/ml, Human, Mouse, Rat Immunoprecipitation, 0.5-2 μg/ml, Human Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of CACO-2 cells using anti-P38 Alpha/MAPK14 antibody. Overlay histogram showing CACO-2 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-P38 Alpha/MAPK14 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IHC analysis of p38 alpha/MAPK14 using anti-p38 alpha/MAPK14 antibody. p38 alpha/MAPK14 was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-p38 alpha/MAPK14 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

IHC analysis of p38 alpha/MAPK14 using anti-p38 alpha/MAPK14 antibody. p38 alpha/MAPK14 was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-p38 alpha/MAPK14 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

IHC analysis of p38 alpha/MAPK14 using anti-p38 alpha/MAPK14 antibody. p38 alpha/MAPK14 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-p38 alpha/MAPK14 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

Immunoprecipitating p38 alpha/MAPK14 in Jurkat whole cell lysate. Western blot analysis of p38 alpha/MAPK14 using anti-p38 alpha/MAPK14 antibody; Lane 1: Jurkat whole cell lysates (30 ug); Lane 2: Rabbit control IgG instead of anti-p38 alpha/MAPK14 antibody in Jurkat whole cell lysate; Lane 3: anti-p38 alpha/MAPK14 antibody (2 µg) + Jurkat whole cell lysate (500 µg). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-p38 alpha/MAPK14 antigen affinity purified polyclonal antibody at a dilution of 0.5 µg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. A specific band was detected for p38 alpha/MAPK14 at approximately 38 kDa. The expected band size for p38 alpha/MAPK14 is at 41 kDa.

Western blot analysis of p38 alpha/MAPK14 using anti-p38 alpha/MAPK14 antibody. Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human Jurkat whole cell lysates, Lane 2: human SW620 whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: human CACO-2 whole cell lysates, Lane 5: rat lung tissue lysates, Lane 6: rat heart tissue lysates, Lane 7: mouse lung tissue lysates, Lane 8: mouse heart tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-p38 alpha/MAPK14 antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate with Tanon 5200 system. A specific band was detected for p38 alpha/MAPK14 at approximately 38 kDa. The expected band size for p38 alpha/MAPK14 is at 41 kDa.