Immunofluorescence analysis of p38 MAPK was done on 70% confluent log phase HeLa cells treated with UV-302nm for 60 minutes and cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with of p38 MAPK Rabbit Polyclonal Antibody (AHO1202) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 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® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing nuclear and cytoplasmic localization. Panel e is untreated cells with no signal. Panel f is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Recombinant human p38-alpha MAPK.|
|Storage buffer||PBS, pH 7.2, with 1% BSA|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||Assay Dependent|
* 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.
AHO1202 has been successfully used in immunofluorescence, and western blot with recombinant proteins and whole cell lysates of human and mouse origins.
AHO1202 specifically recognizes both alpha and beta isoforms of p38, with slight reactivity to p38-gamma.
The protein encoded by this gene is a member of the MAP kinase family. MAP kinases act as an integration point for multiple biochemical signals, and are involved in a wide variety of cellular processes such as proliferation, differentiation, transcription regulation and development. This kinase is activated by various environmental stresses and proinflammatory cytokines. The activation requires its phosphorylation by MAP kinase kinases, or its autophosphorylation triggered by the interaction of MAP3K7IP1/TAB1 protein with this kinase. The substrates of this kinase include transcription regulator ATF2, MEF2C, and MAX, cell cycle regulator CDC25B, and tumor suppressor p53, which suggest the roles of this kinase in stress related transcription and cell cycle regulation, as well as in genotoxic stress response. Four alternatively spliced transcript variants of this gene encoding distinct isoforms have been reported.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Mouse||Not Cited||Growth-factor receptor-bound protein-2 (Grb2) signaling in B cells controls lymphoid follicle organization and germinal center reaction.||Jang IK,Cronshaw DG,Xie LK,Fang G,Zhang J,Oh H,Fu YX,Gu H,Zou Y||Proceedings of the National Academy of Sciences of the United States of America (108:7926)||2011|
|Human||Not Cited||The mitogen-activated protein kinases (MAPK) p38 and JNK are markers of tumor progression in breast carcinoma.||Davidson B,Konstantinovsky S,Kleinberg L,Nguyen MT,Bassarova A,Kvalheim G,Nesland JM,Reich R||Gynecologic oncology (102:453)||2006|
|Human||Not Cited||TNF-alpha promotes a stop signal that inhibits neutrophil polarization and migration via a p38 MAPK pathway.||Lokuta MA,Huttenlocher A||Journal of leukocyte biology (78:210)||2005|
||The mitogen-activated protein kinases (MAPK) p38 and JNK are markers of tumor progression in breast carcinoma.||Davidson B,Konstantinovsky S,Kleinberg L,Nguyen MT,Bassarova A,Kvalheim G,Nesland JM,Reich R||Gynecologic oncology (102:453)||2006|