Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 100 nM Calyculin A (Catalog # 1336) and 20 ng/mL Recombinant Human TNF-a (Catalog # 210-TA) for 10 minutes. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human Phospho-RelA/NF kappa B p65 (S536) Monoclonal Antibody (Catalog # MAB72261) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). 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 1.

Detection of Human Phospho-RelA/NF kappaB p65 (S536) by Simple Western^TM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 100 nM Calyculin A (Catalog # 1336) and 20 ng/mL Recombinant Human TNF-a (Catalog # 210-TA) for 10 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-RelA/NF kappa B p65 (S536) at approximately 66 kDa (as indicated) using 1.0 µg/mL of Rabbit Anti-Human Phospho-RelA/NF kappa B p65 (S536) Monoclonal Antibody (Catalog # MAB72261). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

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 (Catalog # 1336) and Recombinant Human TNF-a (Catalog # 210-TA; upper panel) using Rabbit Anti-Human Phospho-RelA/NF kappa B p65 (S536) Monoclonal Antibody (Catalog # MAB72261) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) 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.

Applications for Human Phospho-RelA/NFkB p65 (S536) Antibody

Application

Recommended Usage

Immunocytochemistry

5-25 µg/mL
Sample: Immersion fixed HT‑29 human colon adenocarcinoma cell line untreated or treated with Calyculin A and Recombinant Human TNF‑ alpha (Catalog # 210-TA)

Simple Western

1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line treated with Calyculin A and Recombinant Human TNF‑ alpha (Catalog # 210-TA)

Western Blot

0.1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line treated with Calyculin A and Recombinant Human TNF‑ alpha (Catalog # 210-TA)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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 kappaB (Nuclear Factor kappa from B cells) complex, that play a fundamental role in inflammatory and immune responses. The NF kappaB complex is composed of a heterodimer of a Rel family member (RelA, c-Rel, RelB) and either NF kappaB1 or NF kappaB2 subunits. RelA and NF kappaB1 are the most common heterodimeric pair. The NF kappaB complex is sequestered in the cytoplasm by inhibitory I kappaB proteins. Upon cellular activation, the ubiquitin-proteosome pathway degrades the I kappaB proteins allowing the NF kappaB complex to translocate to the nucleus and activate gene transcription.

Long Name

v-rel Reticuloendotheliosis Viral Oncogene Homolog A

Alternate Names

NFkB p65, NFKB3, p65RelA

Entrez Gene IDs

5970 (Human)

Gene Symbol

RELA

Additional RelA/NFkB p65 Products

Product Documents for Human Phospho-RelA/NFkB p65 (S536) Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human Phospho-RelA/NFkB p65 (S536) Antibody

For research use only

Citations
Reviews

Human Phospho-RelA/NFkB p65 (S536) Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB72261

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications

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Cited:

Label

Antibody Source

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Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human Phospho-RelA/NFkB p65 (S536) Antibody

Detection of Human Phospho-RelA/NF kappaB p65 (S536) by Western Blot.

Detection of Human Phospho-RelA/NF kappaB p65 (S536) by Simple WesternTM.