Detection of Human Phospho-RelA/NF kappa B p65 (S536) by Western Blot.
Western blot shows lysates of HT‑29 human colon adenocarcinoma cell line and HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 100 nM Calyculin A and 20 ng/mL Recombinant Human TNF‑ alpha (Catalog # 210-TA) for 10 minutes. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Phospho-RelA/NF kappa B p65 (S536) Monoclonal Antibody (Catalog # MAB7226) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-RelA/ NF kappa B p65 (S536) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Phospho-RelA/NFkB p65 (S536) in HT‑29 Human Cell Line.
RelA/NF kappa B p65 phosphorylated at S536 was detected in immersion fixed HT‑29 human colon adenocarcinoma cell line untreated (lower panel) or treated with Calyculin A and Recombinant Human TNF‑ alpha (Catalog # 210-TA; upper panel) using Mouse Anti-Human Phospho-RelA/NF kappa B p65 (S536) Monoclonal Antibody (Catalog # MAB7226) at 2 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and cell surfaces. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: RelA/NFkB p65
RelA belongs to a family of transcription factors (NF kappa B (nuclear factor kappa from B cells) complex) that play a fundamental role in inflammatory and immune responses. The NF kappa B complex is composed of a heterodimer of a Rel family member (RelA, c-Rel, RelB) and either NF kappa B1 or NF kappa B2 subunits. RelA and NF kappa B1 are the most common heterodimeric pair. The NF kappa B complex is sequestered in the cytoplasm by inhibitory I kappa B proteins. Upon cellular activation, the ubiquitin-proteosome pathway degrades the I kappa B proteins allowing the NF kappa B complex to translocate to the nucleus and activate gene transcription.
v-rel Reticuloendotheliosis Viral Oncogene Homolog A
Entrez Gene IDs:
MGC131774; nf kb p65; NFkB p65; NF-kB p65; NFKB3; NFKB3v-rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor ofkappa light polypeptide gene enhancer in B-cells 3 (p65)); Nuclear factor NF-kappa-B p65 subunit; Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3transcription factor p65; p65; p65RelA; rela p65; RelA; v-rel reticuloendotheliosis viral oncogene homolog A (avian); v-rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappalight polypeptide gene enhancer in B-cells 3, p65
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