Human Phospho-FADD (S194) Antibody

Catalog #: MAB7047 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB7047
MAB7047-SP

Detection of Human Phospho-FADD (S194) by Western Blot.

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Human Phospho-FADD (S194) Antibody Summary

Species Reactivity

Human

Specificity

Detects human FADD when phosphorylated on Ser194.

Source

Monoclonal Mouse IgG_2A Clone # 709736

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Phosphopeptide containing the human FADD S194 site

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.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunocytochemistry

8-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human Phospho-FADD (S194) 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 1 µg/mL nocodazole or 4 mM hydroxyurea for 20 hours. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Phospho-FADD (S194) Monoclonal Antibody (Catalog # MAB7047) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for FADD at approximately 28 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

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Phospho-FADD (S194) in HeLa Human Cell Line. FADD phosphorylated at S194 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line stimulated with nocadazole using Mouse Anti-Human Phospho-FADD (S194) Monoclonal Antibody (Catalog # MAB7047) at 10 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 nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL.

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: FADD

Human FADD (Fas-Associating protein with Death Domain), also known as MORT1, is a 25 kDa, 208 amino acid (aa) adaptor protein that mediates signaling of death domain-containing TNF receptor superfamily members and is thus a major regulator of apoptosis. FADD is also implicated in proliferation, cell cycle progression, tumor development, inflammation, innate immunity and autophagy. Phosphorylation on human FADD Ser194 (Ser191 in mouse) is found in T cells arrested at G2/M in the cell cycle; this effect is independent of death domain receptors. Human FADD shares 68% aa identity with mouse and rat FADD.

Long Name

Fas-associated Death Domain Protein

Entrez Gene IDs

8772 (Human)

Alternate Names

FADD; Fas (TNFRSF6)-associated via death domain; FAS-associated death domain protein; FAS-associating death domain-containing protein; GIG3; Growth-inhibiting gene 3 protein; Mediator of receptor induced toxicity; mediator of receptor-induced toxicity; MGC8528; MORT1; MORT1Fas-associating protein with death domain; protein FADD