Human TNF RI/TNFRSF1A Antibody

Name: Human TNF RI/TNFRSF1A Antibody
Brand: R&D Systems
SKU: AB-225-PB
Price: 399 USD
Availability: InStock

R&D Systems | Catalog # AB-225-PB

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Citations (4)

Product Documents

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

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Agonist Activity

Cited:

Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all TNF RI/TNFRSF1A Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human sTNF RI/TNFRSF1A

Specificity

Detects human TNF RI/TNFRSF1A in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human (rh) TNF RII, rhTNF-alpha, recombinant mouse TNF-alpha, recombinant rat TNF-alpha, and rhTNF-beta is observed. Because this antibody preparation is a total IgG fraction, complete monospecificity cannot be assumed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human TNF RI/TNFRSF1A Antibody

Detection of TNF RI/TNFRSF1A by Western Blot

Inhibition of TNF signalling in cells.a Western blots measuring RIP1 ubiquitination following TNFR1 immunoprecipitation from Jurkat cells treated with TNF ± UCB-9260 or etanercept. pNFkB expression was assessed, along with GAPDH for loading control. An isotype control antibody was included as an immunoprecipitation control (lane 5). b TNF (10 pM) or anti-TNFR1 agonist antibody (0.3 mg/mL)-driven NFkB activity was measured following UCB-9260 treatment in Hek-293 cells using a reporter gene assay system. Representative concentration-response curves are shown. Percentage inhibitions for compound dilutions were calculated between a DMSO control and maximum inhibition (by excess biologic anti-TNF, or NFkB inhibitor–TPCA-1). Untransformed data shown in Supplementary Fig. 10c. c UCB-9260 inhibition of TNF (human or mouse) dependent cytotoxicity was measured in mouse L929 cells. Representative concentration-response curves are shown. Percentage inhibition was calculated between unstimulated wells (maximum signal), and wells with TNF, DMSO and actinomycin D (minimum signal). Untransformed data shown in Supplementary Fig. 10d. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31857588), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human TNF RI/TNFRSF1A Antibody

Application

Recommended Usage

Agonist Activity

Measured in a cytotoxicity assay using either L-929 mouse fibrosarcoma cells or A549 human lung carcinoma cells in the presence of the metabolic inhibitor actinomycin D.
The ED₅₀ for this effect is typically 10-15 μg/mL for L-929 cells or 5-10 μg/mL for A549 cells.

Western Blot

1 µg/mL
Sample: Recombinant Human TNF RI/TNFRSF1A (Catalog # 636-R1)

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Reconstitution

Reconstitute at 1 mg/mL in sterile PBS.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it 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: TNF RI/TNFRSF1A

TNF receptor 1 (TNF RI; also called TNF R-p55/p60 and TNFRSF1A) is a type I transmembrane protein member of the TNF receptor superfamily member, designated TNFRSF1A (1, 2). Both TNF RI and TNF RII (TNFRSF1B) are widely expressed and contain four TNF- alpha trimer-binding cysteine-rich domains (CRD) in their extracellular domains (ECD). However, TNF RI is thought to mediate most of the cellular effects of TNF-alpha (3). It is essential for proper development of lymph node germinal centers and Peyer’s patches, and for combating intracellular pathogens such as Listeria (1 - 3). TNF RI is also a receptor for TNF-beta /TNFSF1B (lymphotoxin-alpha ) (4). TNF RI is present on the cell surface as a trimer of 55 kDa subunits (4, 5). TNF-alpha induces sequestering of TNF RI in lipid rafts, where it activates NF kappa B and is cleaved by ADAM-17/TACE (9, 10). Release of the 28 - 34 kDa TNF RI ECD also occurs constitutively and in response to products of pathogens such as LPS, CpG DNA or S. aureus protein A (1, 6 - 8). Full-length TNF RI may also be released in exosome-like vesicles (11). Release helps to resolve inflammatory reactions, since it down-regulates cell surface TNF RI and provides soluble TNF RI to bind TNF-alpha (6, 12, 13). Exclusion from lipid rafts causes endocytosis of TNF RI complexes and induces apoptosis (1). Human TNF RI is a 455 amino acid (aa) protein that contains a 21 aa signal sequence, a 190 aa ECD with a PLAD domain(5) that mediates constitutive trimer formation, followed by the four CRD, a 23 aa transmembrane domain, and a 221 aa cytoplasmic sequence that contains a neutral sphingomyelinase activation domain and a death domain (15). The ECD of human TNF RI shows 80%, 80%, 73%, 69% and 70% aa identity with dog, cat, pig, rat and mouse TNF RI, respectively; it shows 23% aa identity with the ECD of TNF RII.