Anti-ERK1/2 Polyclonal Antibody is a Rabbit antibody targeting ERK1/2. Anti-ERK1/2 Polyclonal Antibody can be used in FCM,IF,IHC-Fr,IHC-P,WB.

IgG

Human,Mouse,Rat (predicted:Chicken,Dog,Pig,Cow,Horse,Rabbit,Sheep,Goat)

1. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (ERK1 + ERK2) Polyclonal Antibody, Unconjugated (TMAB-00633) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
2. Sample:
Brain (Rat) Lysate at 30 μg
Heart (Rat) lysate at 30 μg
Primary: Anti-ERK2/MAPK1 (TMAB-00633) at 1/200 dilution
Secondary: HRP conjugated Goat-Anti-rabbit IgG (secondary antibody) at 1/3000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDa
3. Tissue/cell: human lung carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), 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-ERK2/MAPK1 Polyclonal Antibody, Unconjugated (TMAB-00633) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
4. Blank control: Hep G2 cells (blue). Primary Antibody: Rabbit Anti-ERK1 + ERK2 antibody (TMAB-00633), Dilution: 1 μg 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-Pe (white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice. Primary antibody (TMAB-00633, 1 μg/1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 10% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice.
5. Sample:
Lane 1: A431 (Human) Cell Lysate at 30 μg
Lane 2: MCF-7 (Human) Cell Lysate at 30 μg
Lane 3: Huvec (Human) Cell Lysate at 30 μg
Primary:
Anti-ERK1 + ERK2 (TMAB-00633) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 44/42 kDa
Observed band size: 42/40 kDa
6. Sample:
Lane 1: Cerebrum (Mouse) Lysate at 40 μg
Lane 2: Cerebrum (Rat) Lysate at 40 μg
Lane 3: LympHnode (Mouse) Lysate at 40 μg
Lane 4: LympHnode (Rat) Lysate at 40 μg
Primary:
Anti-ERK1 + ERK2 (TMAB-00633) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 44/42 kDa
Observed band size: 40 kDa

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human ERK2

Human

MAPK3

Mitogen-activated protein kinase 3

Tangles & Tau,MAPK Pathway,Cytoplasmic

Serine/threonine kinase which acts as an essentialcomponent of the MAP kinase signal transduction pathway. MAPK1/ERK2and MAPK3/ERK1 are the 2 MAPKs which play an important role in theMAPK/ERK cascade. They participate also in a signaling cascadeinitiated by activated KIT and KITLG/SCF. Depending on the cellularcontext, the MAPK/ERK cascade mediates diverse biological functionssuch as cell growth, adhesion, survival and differentiation throughthe regulation of transcription, translation, cytoskeletalrearrangements. The MAPK/ERK cascade plays also a role ininitiation and regulation of meiosis, mitosis, and postmitoticfunctions in differentiated cells by phosphorylating a number oftranscription factors. About 160 substrates have already beendiscovered for ERKs. Many of these substrates are localized in thenucleus, and seem to participate in the regulation of transcriptionupon stimulation. However, other substrates are found in thecytosol as well as in other cellular organelles, and those areresponsible for processes such as translation, mitosis andapoptosis. Moreover, the MAPK/ERK cascade is also involved in theregulation of the endosomal dynamics, including lysosome processingand endosome cycling through the perinuclear recycling compartment(PNRC); as well as in the fragmentation of the Golgi apparatusduring mitosis. The substrates include transcription factors (suchas ATF2, BCL6, ELK1, ERF, FOS, HSF4 or SPZ1), cytoskeletal elements(such as CANX, CTTN, GJA1, MAP2, MAPT, PXN, SORBS3 or STMN1),regulators of apoptosis (such as BAD, BTG2, CASP9, DAPK1, IER3,MCL1 or PPARG), regulators of translation (such as EIF4EBP1) and avariety of other signaling-related molecules (like ARHGEF2, DCC,FRS2 or GRB10). Protein kinases (such as RAF1, RPS6KA1/RSK1,RPS6KA3/RSK2, RPS6KA2/RSK3, RPS6KA6/RSK4, SYK, MKNK1/MNK1,MKNK2/MNK2, RPS6KA5/MSK1, RPS6KA4/MSK2, MAPKAPK3 or MAPKAPK5) andphosphatases (such as DUSP1, DUSP4, DUSP6 or DUSP16) are othersubstrates which enable the propagation the MAPK/ERK signal toadditional cytosolic and nuclear targets, thereby extending thespecificity of the cascade. Mediates phosphorylation of TPR inrespons to EGF stimulation. May play a role in the spindle assemblycheckpoint. Phosphorylates PML and promotes its interaction withPIN1, leading to PML degradation (By similarity).
Acts as a transcriptional repressor. Binds to a[GC]AAA[GC] consensus sequence. Repress the expression ofinterferon gamma-induced genes. Seems to bind to the promoter ofCCL5, DMP1, IFIH1, IFITM1, IRF7, IRF9, LAMP3, OAS1, OAS2, OAS3 andSTAT1. Transcriptional activity is independent of kinase activity.