Human phospho-RelA/NFkB p65 (S536) Antibody (1091B)

Name: Human phospho-RelA/NFkB p65 (S536) Antibody (1091B)
Brand: R&D Systems
SKU: MAB72261
Price: 169 USD
Availability: InStock

R&D Systems | Catalog # MAB72261

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Citations (13)

Product Documents

Recombinant Monoclonal Antibody.

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Immunocytochemistry, Simple Western

Cited:

Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 1091B

View all RelA/NFkB p65 Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing the RelA/NF kappa B p65 S536 site

Specificity

Detects human RelA/NF kappa B p65 when phosphorylated at S536.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human phospho-RelA/NFkB p65 (S536) Antibody (1091B)

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

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 kappa B 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 (1091B)

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

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.