WB
Western blot analysis of Phospho-PKC alpha/beta II (T638/T641) on different lysates with Rabbit anti-Phospho-PKC alpha/beta II (T638/T641) antibody at 1/25,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 14 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/25,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue untreated / treated with λpp with Rabbit anti-Phospho-PKC alpha/beta II (T638/T641) antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of HeLa cells untreated / treated with λpp labeling Phospho-PKC alpha/beta II (T638/T641) with Rabbit anti-Phospho-PKC alpha/beta II (T638/T641) antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-PKC alpha/beta II (T638/T641) antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin ( red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of NIH/3T3 cells untreated (left) / treated with λpp (right) labeling Phospho-PKC alpha/beta II (T638/T641). Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).DB
Dot blot analysis of Phospho-PKC alpha/beta II (T638/T641) on different peptides with Rabbit anti-Phospho-PKC alpha/beta II (T638/T641) antibody at 1/5,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution for 1 hour at room temperature. Proteins loading: 100ng, 25ng, 5ng; Blocking and dilution buffer: 5% NFDM/TBST; Exposure time: 3 seconds .| Product Name | Phospho-PKC alpha/beta II (T638/T641) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic phospho-peptide corresponding to residues surrounding Thr638 of Human PKC alpha aa 622-667 / 672. |
| Modification | p-T638/T641 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | DBFCICC/IFIHCWB |
| WB:1:25000 IHC:1:200 ICC/IF:1:100 FC:1:1000 DB:1:5000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | PRKCA |
|---|---|
| Gene Synonyms | AAG6 PKCA PKCI+/- PKCalpha PKC-alpha |
| Gene Full Name | protein kinase C alpha |
| Gene Summary | Protein kinase C (PKC) is a family of serine- and threonine-specific protein kinases that can be activated by calcium and the second messenger diacylglycerol. PKC family members phosphorylate a wide variety of protein targets and are known to be involved in diverse cellular signaling pathways. PKC family members also serve as major receptors for phorbol esters, a class of tumor promoters. Each member of the PKC family has a specific expression profile and is believed to play a distinct role in cells. The protein encoded by this gene is one of the PKC family members. This kinase has been reported to play roles in many different cellular processes, such as cell adhesion, cell transformation, cell cycle checkpoint, and cell volume control. Knockout studies in mice suggest that this kinase may be a fundamental regulator of cardiac contractility and Ca(2+) handling in myocytes. [provided by RefSeq, Jul 2008] |
| Alternative Names | AAG6 antibody Aging associated gene 6 antibody aPKC antibody KPCA_HUMAN antibody PKC alpha antibody PKC-A antibody PKC-alpha antibody PKCA antibody PRKACA antibody PRKCA antibody AAG6 antibody Aging associated gene 6 antibody aPKC antibody KPCA_HUMAN antibody PKC alpha antibody PKC-A antibody PKC-alpha antibody PKCA antibody PRKACA antibody PRKCA antibody Protein Kinase C alpha antibody Protein kinase C alpha type antibody KPCB_HUMAN antibody PKC Beta antibody PKC-B antibody PKC-beta antibody PKCB antibody Prkcb antibody PRKCB II antibody PRKCB2 antibody Protein kinase C beta antibody Protein kinase C beta type antibody |
| Molecular Weight(MW) | 77 kDa |
| Cellular Localization | Cytoplasm, Cell membrane, Mitochondrion membrane, Nucleus. |
WB
Western blot analysis of Phospho-PKC alpha/beta II (T638/T641) on different lysates with Rabbit anti-Phospho-PKC alpha/beta II (T638/T641) antibody at 1/25,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 14 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/25,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue untreated / treated with λpp with Rabbit anti-Phospho-PKC alpha/beta II (T638/T641) antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of HeLa cells untreated / treated with λpp labeling Phospho-PKC alpha/beta II (T638/T641) with Rabbit anti-Phospho-PKC alpha/beta II (T638/T641) antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-PKC alpha/beta II (T638/T641) antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin ( red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of NIH/3T3 cells untreated (left) / treated with λpp (right) labeling Phospho-PKC alpha/beta II (T638/T641). Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
DB
Dot blot analysis of Phospho-PKC alpha/beta II (T638/T641) on different peptides with Rabbit anti-Phospho-PKC alpha/beta II (T638/T641) antibody at 1/5,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution for 1 hour at room temperature. Proteins loading: 100ng, 25ng, 5ng; Blocking and dilution buffer: 5% NFDM/TBST; Exposure time: 3 seconds .| Application Notes | WB:1:25000 IHC:1:200 ICC/IF:1:100 FC:1:1000 DB:1:5000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
