Anti-ERK1/2 Antibody (6Q27) is a Rabbit antibody targeting ERK1/2. Anti-ERK1/2 Antibody (6Q27) can be used in FCM,ICC/IF,IF,IHC-Fr,IHC-P,WB.

IgG

6Q27

Human,Mouse,Rat

1. Tissue/cell: A549 cell;4% Paraformaldehyde-fixed;Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (ERK1/2) monoclonal Antibody, Unconjugated (TMAB-00637) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
2. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-ERK1/2 antibody (TMAB-00637)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 1 μg/test.
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.
3. Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded Human Cervical Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded Human Breast; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
10. Paraformaldehyde-fixed, paraffin embedded Human Placenta; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
11. Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
12. Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
13. Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
14. Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
15. Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
16. Paraformaldehyde-fixed, paraffin embedded Human Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
17. Paraformaldehyde-fixed, paraffin embedded Human Glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
18. Paraformaldehyde-fixed, paraffin embedded Human Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
19. Paraformaldehyde-fixed, paraffin embedded Human Pancreas; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated (TMAB-00637) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
20. 25 μg total protein per Lane of various lysates probed with ERK1/2 monoclonal antibody, unconjugated (TMAB-00637) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at RT for 60 min.
21. 4% Paraformaldehyde-fixed Hela (H) cell; Triton X-100 at RT for 20 min; Antibody incubation with (ERK1/2) monoclonal Antibody, unconjugated (TMAB-00637) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green) at 37°C for 90 min, DAPI (blue) was used to stain the cell nucleus. PBS instead of the primary antibody was used as the blank control.
22. The Hela (H) 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 then were incubated in 5% BSA to block non-specific protein-protein interactions (30 min at room temperature). Primary Antibody (green): Rabbit Anti-ERK1/2 antibody (TMAB-00637): 1 μg/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-BF488: 1 μg/test. Isotype Control (orange): Rabbit IgG. Blank control (black): PBS.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:50-200; IF: 1:100-500; FCM: 1ug/Test

Monoclonal

KLH conjugated synthetic peptide: human ERK1/2

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.