Phospho-NF-κB p65 (S529) Recombinant Rabbit Monoclonal Antibody [SP07-00]
cat.: ET1604-27
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IHC-P, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | SP07-00 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 60 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser529 of Human NF-kB p65. |
| Positive control: | HeLa treated with 100nM Calyculin A and 20ng/mL TNF-α for 15 minutes cell lysate, human lung tissue, human lung squamous cell carcinoma tissue, Daudi. |
| Subcellular location: | Nucleus, Cytoplasm. |
| Recommended Dilutions:
WB IHC-P FC IP |
1:1,000 1:1,000 1:50 Use at an assay dependent concentration. |
| Uniprot #: | SwissProt: Q04206 Human |
| Alternative names: | Avian reticuloendotheliosis viral (v rel) oncogene homolog A MGC131774 NF kappa B p65delta3 NFKB3 Nuclear Factor NF Kappa B p65 Subunit Nuclear factor NF-kappa-B p65 subunit Nuclear factor of kappa light polypeptide gene enhancer in B cells 3 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 OTTHUMP00000233473 OTTHUMP00000233474 OTTHUMP00000233475 OTTHUMP00000233476 OTTHUMP00000233900 p65 p65 NF kappaB p65 NFkB relA TF65_HUMAN Transcription factor p65 v rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor of kappa light polypeptide gene enhancer in B cells 3 (p65)) V rel avian reticuloendotheliosis viral oncogene homolog A v rel reticuloendotheliosis viral oncogene homolog A (avian) V rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappa light polypeptide gene enhancer in B cells 3, p65 |
Images
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Fig1:
Western blot analysis of Phospho-NF-κB p65 (S529) on different lysates with Rabbit anti-Phospho-NF-κB p65 (S529) antibody (ET1604-27) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 100nM Calyculin A and 20ng/mL TNF-α for 15 minutes cell lysate Lane 3: HeLa treated with 100nM Calyculin A and 20ng/mL TNF-α for 15 minutes cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 60 kDa Observed band size: 65 kDa Exposure time: 43 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 (ET1604-27) at 1/1,000 dilution was used in 5% NFDM/TBST 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:
Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-Phospho-NF-κB p65 (S529) antibody (ET1604-27) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1604-27) at 1/1,000 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. |
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Fig3:
Immunohistochemical analysis of paraffin-embedded human lung squamous cell carcinoma tissue with Rabbit anti-Phospho-NF-κB p65 (S529) antibody (ET1604-27) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1604-27) at 1/1,000 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. |
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Fig4: Flow cytometric analysis of Phospho-NF-κB p65 (S529) was done on Daudi cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1604-27, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.