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Anti-RELA Antibody 27F9.G4
Also for RELA (NM_001145138)
|NFkB p65 (Rel A) peptide corresponding to a region near the C-terminus of the human protein conjugated to Keyhole Limpet Hemocyanin (KLH).|
|WB, IHC, IF
||ELISA: 1:50,000-1:100,000, WB: 1:1,000-1:5,000, IHC: 1:200-1:600, IF: 1:5,000
|0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2|
|NFkappaB was originally identified as a factor that binds to the immunoglobulin kappa light chain enhancer in B cells. Other identified subunits include p52 (NFKB2), c-Rel, and RelB. The p65, cRel, and RelB subunits are responsible for transactivation. The p50 and p52 subunits possess DNA binding activity but limited ability to transactivate. p52 has been reported to form transcriptionally active heterodimers with the NFkappaB subunit p65, similar to p50/p65 heterodimers. The heterodimers of p52/p65 and p50/p65 are regulated by physical inactivation in the cytoplasm by IkappaBalpha. Cell Biology, Nuclear Signaling, Neuroscience and Signal Transduction Research.
|Homo sapiens v-rel avian reticuloendotheliosis viral oncogene homolog A (RELA), transcript variant 2|
Delta-Notch Signaling Pathway
MAPK signaling pathway
Toll-like receptor signaling pathway
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anti NFKB p65 (Rel A) monoclonal antibody 200-301-065 was used to detect ~65 kD band (red arrow) in HeLa whole cell lysate. Lysate was run on 4-20% gradient gel transferred under standard conditions and blocked in 1% BSA-TTBS 30 min RT. Blot was probed with monoclonal anti p65 1:1000 in 1% BSA-TBS-T o/n 4°C and detected with HRP conjugated Rb-anti Mouse antibody 1:40,000 in MB-070 30 min RT.
monoclonal anti p65 200-301-065 was used to detect p65 by Western blot. Samples were prepared in RIPA lysis buffer, boiled with NuPage 4x LDS Sample Buffer and run on NuPage 4-12% Bis-Tris Gels. Blot was incubated with primary antibody at a dilution of 1:500 and detected with HRP conjugated anti mouse antibody at a dilution of 1:10000. Image provided courtesy of Dr. Al Baldwin, University of North Carolina, Chapel Hill.
Antibody 200-301-065 has been tested in immunohistochemistry, analyzed by an anatomic pathologist and validated for use in IHC applications against formalin-fixed, paraffin-embedded human tissues. showed moderate to strong staining within many tissues, including epithelium of the breast, small intestine, kidney, pancreas, prostate, skin, placenta, and uterus, as well as within neurons and lymphoid tissues such as spleen, thymus, and tonsil. The antibody produced an excellent signal with almost no background staining at a concentration of 2.5 ug/ml. The image displayed shows specific staining in colon carcinoma as the precipitated red signal, with a hematoxylin purple nuclear counterstain. Image provided courtesy of LifeSpan Biosciences, Seattle, WA
Monoclonal anti NFKB p65 (Rel A) antibody was used to detect p65 by immunofluorescence at a dilution of 1:5000. Hela cells were grown to sub-confluent on 18 mm2 glass coverslips #1.5. Cells were either unstimulated (A), or stimulated (B) with 50 ng/ml of TNF alpha for 30 min prior fixation. Cells were then fixed in methanol and blocked with 10% normal goat serum (NGS), in PBS, and TritonX 0.2% (Tx) and incubated for 1 hr at RT with primary ab, counterstained with DAPI and washed in PBS/NGS/Tx. Cells were incubated for 1 hr at RT with Atto 425 conjugated anti mouse secondary antibody for STED CW imaging. Data was collected on a STED-CW TCS-SP5 Confocal system equipped with a DFC 350FX camera allowing sequential acquisition in widefiled, confocal and STED CW imaging on the same system.