Immunohistochemistry analysis of p38 MAPK [pTpY180/182] showing staining in the cytoplasm of paraffin-embedded human colon carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a p38 MAPK [pTpY180/182] monoclonal antibody (701057) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Phosphopeptide corresponding to amino acids 176–186 of human p38 MAPK|
|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 *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||0.1-2 ug/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.
|Miscellaneous PubMed (MISC)||See 1 publications below|
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.
p38 Mitogen-activated protein kinase (MAPK) belongs to the mitogen-activated protein kinase family, which responds to stress stimuli. The p38 MAPK subfamily plays an important role in cytokine production and the stress response. Members are activated by UV irradiation, heat shock, high osmotic stress, protein synthesis inhibitors, lipopolysaccharides, proinflammatory cytokines (e.g., IL-1 and TNF-alpha) and certain mitogens. There are four p38 kinases ( alpha, beta, lambda, and delta), with p38 alpha being well characterized, and expressed in most cell types. Additional functions for p38 MAPK include inhibition of cell cycle progression, development, and differentiation. An important target of p38 MAPK is the tumor suppressor protein p53, whose activation is positively regulated by p38. MKK3, MKK6 and SEK phosphorylate p38 MAPK at threonine 180 and tyrosine 182. Phosphorylation of these residues activates p38 MAPK, which in turn activates and phosphorylates ATF1, ATF2 and ATF6.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Possible Mechanisms of Di(2-ethylhexyl) Phthalate-Induced MMP-2 and MMP-9 Expression in A7r5 Rat Vascular Smooth Muscle Cells.
701057 was used in western blot to test if DEHP affects MMP-2 or MMP-9 expression in vascular smooth muscle cells
|Shih MF,Pan KH,Cherng JY||International journal of molecular sciences (16:28800)||2015|