Immunofluorescent analysis of PKC alpha (green) in SH-SY5Y and MCF7 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a PKC alpha monoclonal antibody (Product # MA1-157) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502). F-Actin (red) was stained with DyLight-554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Bovine, Dog, Human, Mouse, Non-human primate, Rabbit, Rat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Purified bovine brain Protein Kinase C alpha.|
|Storage buffer||PBS with 30% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:50 - 1:100|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Immunoprecipitation (IP)||5 µg|
|Western Blot (WB)||1:1000 - 1:2000|
* 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.
|Immunohistochemistry (IHC)||See 1 publications below|
Western blot analysis of MA1-157 detects an ~80 kDa protein in human, bovine, non-human primate, mouse, rat, dog and rabbit cells. In Jurkat, rat brain and MDCK cell lysates, a weak unknown band at ~140 kDa is also detected. MA1-157 shows specificity to PKC alpha and is non-reactive to other PKC family members.
Members of the Protein kinase C (PKC) family of serine/threonine protein kinases are grouped by their activation mechanism. Classical or conventional PKCs (PKC alpha-, betaI- , betaII- and gamma-) are activated by
phosphatidylserine in a calcium dependent manner and can bind diacylglycerol. The Ca2+ insensitive novel PKCs (PKCs epsilon-, delta-, theta- and eta) are also activated by diacylglycerol and phosphatidylserine. The atypical PKCs (PKCs iota- and zeta-) are insensitive to Ca2+, DAG and phorbolesters. All PKCs isoforms consist of a highly conserved catalytic domain connected to a regulatory domain via a hinge region. PKCs are involved in positive and negative regulation of cell proliferation, apoptosis, differentiation, migration and adhesion, tumorigenesis, cardiac hypertrophy, angiogenesis, platelet function and inflammation, by directly phosphorylating targets such as RAF1, BCL2, CSPG4, TNNT2/CTNT, or activating signaling cascades involving MAPK1/3 (ERK1/2) and RAP1GAP.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Localization of Rod Bipolar Cells in the Mammalian Retina Using an Antibody Against the ¿1c L-type Ca(2+) Channel.
MA1-157 was used in immunohistochemistry to test if the alpha1c subunit of L-type voltage-gated calcium channel colocalizes with protein kinase C alpha in the rod bipolar cells in mouse, hamster, rabbit, and dog retinas
|Huh YJ,Choi JS,Jeon CJ||Acta histochemica et cytochemica (48:47)||2015|