WB
Western blot analysis using OCT-4 antibody against Hela(1),Hepg2(2),Mcf-7(3),A549(4),Cos7(5),A431(6),K562(7) cell 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 lung cancer tissues using OCT-4 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 ddH₂O 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.ICC/IF
Immunofluorescence analysis of U251 cells using OCT-4 antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 20 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Rabbit IgG(H&L) (Green,OM643486), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutes.IHC
Immunohistochemical analysis of paraffin-embedded Human breast cancer tissues using OCT-4 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 ddH₂O 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 colon cancer tissues using OCT-4 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 ddH₂O 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 lung cancer tissues using OCT-4 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 ddH₂O 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-OCT-4 antibody,pbs only |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Polypeptide |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ELISAICC/IFIHCWB |
| WB:1:1000-1:2000 IHC:1:200-1:2000 ICC/IF:1:100-1:500 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | POU5F1 |
|---|---|
| Gene Synonyms | NF-A3 Oct-3 Oct-3/4 Oct-4 OCT3 Oct3/4 OCT4 OCT4Borf1 OTF-3 Otf-4 OTF3 Otf3-rs7 Otf3g OTF4 |
| Gene Full Name | POU class 5 homeobox 1 |
| Gene Summary | This gene encodes a transcription factor containing a POU homeodomain that plays a key role in embryonic development and stem cell pluripotency. Aberrant expression of this gene in adult tissues is associated with tumorigenesis. This gene can participate in a translocation with the Ewing's sarcoma gene on chromosome 21, which also leads to tumor formation. Alternative splicing, as well as usage of alternative AUG and non-AUG translation initiation codons, results in multiple isoforms. One of the AUG start codons is polymorphic in human populations. Related pseudogenes have been identified on chromosomes 1, 3, 8, 10, and 12. [provided by RefSeq, Oct 2013] |
| Alternative Names | 3-Oct 4-Oct DADB-104B20.2 gp-9 HGNC:9221 MGC22487 non-spliced variant Oct-3/4 OCT4-B1 OCT4B-variant 2 octamer-binding protein 3 octamer-binding protein 4 octamer-binding transcription factor 3 octamer-binding transcription factor 4 alternative OTTHUMP00000221150 OTTHUMP00000221151 POU class 5 homeobox 1 POU domain transcription factor OCT4 pou-2 POU-type homeodomain-containing DNA-binding protein pou2 transcriptional regulator |
| Molecular Weight(MW) | 39 kDa |
| Cellular Localization | Cytoplasm, Nucleus |
| SwissProt ID | Q01860 |
|---|
WB
Western blot analysis using OCT-4 antibody against Hela(1),Hepg2(2),Mcf-7(3),A549(4),Cos7(5),A431(6),K562(7) cell 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 lung cancer tissues using OCT-4 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 ddH₂O 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.
ICC/IF
Immunofluorescence analysis of U251 cells using OCT-4 antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 20 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Rabbit IgG(H&L) (Green,OM643486), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutes.
IHC
Immunohistochemical analysis of paraffin-embedded Human breast cancer tissues using OCT-4 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 ddH₂O 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 colon cancer tissues using OCT-4 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 ddH₂O 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 lung cancer tissues using OCT-4 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 ddH₂O 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 ICC/IF:1:100-1:500 |
|---|
| 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 | PBS ONLY |
