Phospho-PKC alpha/beta II (T638/T641) Recombinant Rabbit Monoclonal Antibody [PSH15-91]
cat.: HA723796
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, Dot Blot |
| Clonality: | Monoclonal |
| Clone number: | PSH15-91 |
| Form: | Liquid |
| Storage condition: | 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: | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 77 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Thr638 of Human PKC alpha aa 622-667 / 672. |
| Positive control: | HEK-293 cell lysate, HeLa cell lysate, C6 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, NIH/3T3 cell lysate, NIH/3T3 starved overnight then treated with 200nM TPA for 4 hours cell lysate, HeLa, NIH/3T3, C6. |
| Subcellular location: | Cytoplasm, Cell membrane, Mitochondrion membrane, Nucleus. |
| Recommended Dilutions:
WB IF-Cell Dot Blot |
1:25,000 1:100 1:5,000 |
| Uniprot #: | SwissProt: P17252 Human | P05771-2 Human | P20444 Mouse | P68404-2 Mouse | P05696 Rat | P68403-2 Rat |
| Alternative names: | AAG6 Aging associated gene 6 aPKC KPCA_HUMAN PKC alpha PKC-A PKC-alpha PKCA PRKACA PRKCA Protein Kinase C alpha Protein kinase C alpha type KPCB_HUMAN PKC Beta PKC-B PKC-beta PKCB Prkcb PRKCB II PRKCB2 Protein kinase C beta Protein kinase C beta type |
Images
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Fig1:
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 (HA723796) at 1/25,000 dilution. Lane 1: HEK-293 cell lysate Lane 2: HeLa cell lysate Lane 3: C6 cell lysate Lane 4: Mouse brain tissue lysate Lane 5: Rat brain tissue lysate Lane 6: NIH/3T3 cell lysate Lane 7: NIH/3T3 starved overnight then treated with 200nM TPA for 4 hours cell lysate Lane 8: NIH/3T3 starved overnight then treated with 200nM TPA for 4 hours cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 77 kDa Observed band size: 77 kDa Exposure time: 14 seconds; ECL: K1801; 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 (HA723796) at 1/25,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
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 (HA723796) at 1/5,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution for 1 hour at room temperature. Lane 1: Phospho-PKC alpha (T638) peptide (positive) Lane 2: Phospho-PKC beta II (T641) peptide (positive) Lane 3: Phospho-PKC beta I (T642) peptide (negative) Lane 4: Unmodified PKC alpha peptide (negative) Proteins loading: 100ng, 25ng, 5ng; Blocking and dilution buffer: 5% NFDM/TBST; Exposure time: 3 seconds; ECL: K1801. |
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Fig3:
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 (HA723796) 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 (HA723796) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Immunocytochemistry analysis of NIH/3T3 cells untreated / treated with λpp labeling Phospho-PKC alpha/beta II (T638/T641) with Rabbit anti-Phospho-PKC alpha/beta II (T638/T641) antibody (HA723796) 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 (HA723796) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig5:
Immunocytochemistry analysis of C6 cells untreated / treated with λpp labeling Phospho-PKC alpha/beta II (T638/T641) with Rabbit anti-Phospho-PKC alpha/beta II (T638/T641) antibody (HA723796) 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 (HA723796) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.