Immunofluorescence analysis of ERK 1/2 [pTpY185/187] was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton™ X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with ABfinity™ ERK 1/2 [pTpY185/187] Recombinant Rabbit Monoclonal Antibody (700012) at 2 µg-4 µg in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 Phalloidin (A12381). Panel d is a merged image showing ERK 1/2 [pTpY185/187] in nucleus. Panel e shows no primary antibody control. Panel f shows competition with ERK 1/2 [pTpY185/187] peptide. The images were captured at 20X magnification.
|Tested species reactivity||Dog, Human, Rat|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A peptide corresponding to amino acids 198-208 and 181-191 of P27361 and P28482, respectively.|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1 ul|
|Flow Cytometry (Flow)||2-4µg/10^6 cells|
|Immunohistochemistry (Paraffin) (IHC (P))||5 ug/ml|
|Western Blot (WB)||0.5-1µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 2 publications below|
700012 has successfully been used in western blot to detect phospho-ERK1/2 in TPA-treated HeLa, NRK and MDCK cell lysates.
This antibody is predicted to react with bovine, chicken, chimpanzee, mouse, rat, Xenopus and zebrafish based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
Mitogen activated protein kinase (MAPK) belongs to a family of serine/threonine kinases that are activated upon exposure of cells to a variety of stimuli including mitogens, hormones, growth factors, cytokines and bioactive peptides. There are two closely related isoforms of MAPK, with apparent molecular weights of 44 kDa and 42 kDa. They are abundantly expressed in neurons of the mature central nervous system. Activated MAPK plays a role in cell division and differentiation, and regulation of the cytoskeleton.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
ANKHD1 silencing inhibits Stathmin 1 activity, cell proliferation and migration of leukemia cells.
700012 was used in western blot to identify binding partners ANKHD1 in leukemia cells
|Machado-Neto JA,Lazarini M,Favaro P,de Melo Campos P,Scopim-Ribeiro R,Franchi Junior GC,Nowill AE,Lima PR,Costa FF,Benichou S,Olalla Saad ST,Traina F||Biochimica et biophysica acta (1853:583)||2015|
Multidimensional profiling of CSF1R screening hits and inhibitors: assessing cellular activity, target residence time, and selectivity in a higher throughput way.
700012 was used in western blot to develop medium throughput assays to characterize CSF1R HTS hits and aid the selection of the best compounds for lead progression
|Uitdehaag JC,Sünnen CM,van Doornmalen AM,de Rouw N,Oubrie A,Azevedo R,Ziebell M,Nickbarg E,Karstens WJ,Ruygrok S||Journal of biomolecular screening (16:1007)||2011|