WB
Western blot analysis using p66shc antibody against B16(1) cell lysate and Mouse intestine(2),Mouse liver(3) lysate. 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S.IHC
Immunohistochemical analysis of paraffin-embedded Human breast cancer tissues using p66shc antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH?O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissues using p66shc antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH?O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.ELISA
Coat the plate with immunizing antigen.Blue line： Immune serum; Orange line: Pre-immune serum.| Product Name | Anti-p66shc antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | SHC-transforming protein 1 (UniProt: P29353; also known as SHC-transforming protein 3, SHC-transforming protein A, Src homology 2 domain-containing-transforming protein C1, SH2 domain protein C1) is encoded by the SHC1 (also known as SHC, SHCA) gene (Gene ID: 6464) in human. p66Shc is a splice variant of p52Shc/p46Shc with a typical domain organization that is shared by all members of the Shc family of adaptor proteins. It has a phosphotyrosine binding domain, a collagen homology 1 region, and an SH2 domain. p66Shc is shown to be a downstream target of p53 and activated p53 induces its up-regulation by increasing its stability. p66Shc is suggested to be a genetic determinant of aging in mammals as its homozygous deletion is reported to delay senescence and prolong life span in mice. It regulates the mitochondrial pathway of apoptosis by inducing mitochondrial damage after dissociation from an inhibitory protein complex. Under conditions of excessive reactive oxygen species (ROS), p66Shc is phosphorylated on serine 36 by protein kinase C and is targeted to the mitochondrial intermembrane space where it serves as a redox enzyme by oxidizing cytochrome c and reducing oxygen to form ROS. In the absence of p66Shc, release of cytochrome c from mitochondria is impaired and apoptosome can only be activated if purified cytochrome c is supplied. p66Shc induces the collapse of the mitochondrial trans-membrane potential after oxidative stress. p66Shc is also shown to have a neuroprotective role in a murine model of multiple sclerosis. p66Shc knockout (p66-KO) mice develop typical signs of experimental autoimmune encephalomyelitis (EAE). (Ref.: Su, KG., et al. (2012). Eur. J. Neurosci. 35; 562-571; Orsini, F., et al. (2004). J. Biol. Chem. 279(24); 25689-25695). |
| Immunogen | Polypeptide |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ELISAIHCWB |
| WB:1:1000-1:2000 IHC:1:200-1:2000 ELISA:1:1000-1:10000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | SHC1 |
|---|---|
| Gene Synonyms | p66 P66shc SHC SHCA |
| Gene Full Name | SHC adaptor protein 1 |
| Gene Summary | This gene encodes three main isoforms that differ in activities and subcellular location. While all three are adapter proteins in signal transduction pathways, the longest (p66Shc) may be involved in regulating life span and the effects of reactive oxygen species. The other two isoforms, p52Shc and p46Shc, link activated receptor tyrosine kinases to the Ras pathway by recruitment of the GRB2/SOS complex. p66Shc is not involved in Ras activation. Unlike the other two isoforms, p46Shc is targeted to the mitochondrial matrix. Several transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Feb 2011] |
| Alternative Names | FLJ26504 OTTHUMP00000035472 RP11-307C12.1 SH2 domain protein C1 SHC (Src homology 2 domain containing) transforming protein 1 SHC-transforming protein 3 SHC-transforming protein A src homology 2 domain-containing-transforming protein C1 |
| Molecular Weight(MW) | 63kDa |
| SwissProt ID | P29353 |
|---|
WB
Western blot analysis using p66shc antibody against B16(1) cell lysate and Mouse intestine(2),Mouse liver(3) lysate. 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S.
IHC
Immunohistochemical analysis of paraffin-embedded Human breast cancer tissues using p66shc antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH?O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissues using p66shc antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH?O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
ELISA
Coat the plate with immunizing antigen.Blue line： Immune serum; Orange line: Pre-immune serum.| Application Notes | WB:1:1000-1:2000 IHC:1:200-1:2000 ELISA:1:1000-1:10000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. |
| Storage Buffer | Purified antibody in PBS with 0.05% sodium azide,40% Glycerol. |
