MAPK4 Rabbit Polyclonal Antibody

• Catalog Number: orb5182
• Category: Antibodies
• Description: MAPK4 Rabbit Polyclonal Antibody
• Target: MAPK4
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Predicted Reactivity: Bovine, Canine, Equine, Porcine, Rabbit
• Form/Appearance: Liquid
• Concentration: 1mg/ml
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Immunogen: KLH conjugated synthetic peptide derived from human ERK5 (401-500/587aa)
• UniProt ID: P31152
• RRID: AB_10930938
• MW: 66 kDa
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1ug/Test
• Antibody Type: Primary Antibody
• 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: ERK-4; ERK3; Erk3 related; ERK4; Extracellular sig
• Note: For research use only

Blank control (black line): Hela. Primary Antibody (green line): Rabbit Anti-MAPK4 antibody (orb5182), dilution: 1 ug/Test, Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488, dilution: 0.5 ug/Test. Isotype control (orange line): Normal Rabbit IgG, Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C, 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 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (MAPK4) Polyclonal Antibody, Unconjugated (orb5182) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (MAPK4) Polyclonal Antibody, Unconjugated (orb5182) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (MAPK4) Polyclonal Antibody, Unconjugated (orb5182) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat brain tissue), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (ERK4) Polyclonal Antibody, Unconjugated (orb5182) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (MAPK4) Polyclonal Antibody, Unconjugated (orb5182) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat cerebellum), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (MAPK4) Polyclonal Antibody, Unconjugated (orb5182) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat kidney tissue), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (ERK4) Polyclonal Antibody, Unconjugated (orb5182) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Sample: Lane 1: Mouse Cerebellum tissue lysates, Lane 2: Rat Cerebellum tissue lysates, Lane 3: Human Raji cell lysates, Lane 4: Human HeLa cell lysates, Lane 5: Human SH-SY5Y cell lysates, Lane 6: Human U251 cell lysates, Lane 7: Human 293T cell lysates, Primary: Anti-MAPK4 (orb5182) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 66 kDa, Observed band size: 60 kDa.

Tissue/Cell: mouse brain tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-ERK4 Polyclonal Antibody, Unconjugated (orb5182) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Tissue/Cell: rat brain tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-ERK4 Polyclonal Antibody, Unconjugated (orb5182) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.