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Rat TNF alpha Recombinant Rabbit Monoclonal Antibody [PSH04-70] - BSA and Azide free (Capture)

Usd: 649

Specification

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Catalog# HA722170

Rat TNF alpha Recombinant Rabbit Monoclonal Antibody [PSH04-70] - BSA and Azide free (Capture)

Application

  • ELISA(Cap)

Reactivity

  • Rat

Pair: Capture
Pair: Detector
Pair: Protein
Range

  • 12.3-3,000 pg/mL

Conjugation

  • unconjugated

Overview

Product Name

Rat TNF alpha Recombinant Rabbit Monoclonal Antibody [PSH04-70] - BSA and Azide free (Capture)

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Rat TNF alpha aa 80-235 (P16599).

Species Reactivity

Rat

Validated Applications

ELISA(Cap)

Molecular Weight

Predicted band size: 26 kDa

Positive Control

Recombinant Rat TNF-α protein (HA210538).

Conjugation

unconjugated

Clone Number

PSH04-70

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • ELISA(Cap)

  • Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH04-71] to Rat TNF alpha (Detector) (HA722171) and recombinant standard Rat TNF alpha (HA210538). The reference range value is 12.3-3,000 pg/mL.

Target

Function

Tumor necrosis factor (TNF, cachexin, or cachectin; formerly known as tumor necrosis factor alpha or TNF-α) is an adipokine and a cytokine. TNF is a member of the TNF superfamily, which consists of various transmembrane proteins with a homologous TNF domain. As an adipokine, TNF promotes insulin resistance, and is associated with obesity-induced type 2 diabetes. As a cytokine, TNF is used by the immune system for cell signaling. If macrophages (certain white blood cells) detect an infection, they release TNF to alert other immune system cells as part of an inflammatory response. TNF signaling occurs through two receptors: TNFR1 and TNFR2. TNFR1 is constituitively expressed on most cell types, whereas TNFR2 is restricted primarily to endothelial, epithelial, and subsets of immune cells. TNFR1 signaling tends to be pro-inflammatory and apoptotic, whereas TNFR2 signaling is anti-inflammatory and promotes cell proliferation. Suppression of TNFR1 signaling has been important for treatment of autoimmune diseases, whereas TNFR2 signaling promotes wound healing.

Background References

1. Jang DI et al. The Role of Tumor Necrosis Factor Alpha (TNF-α) in Autoimmune Disease and Current TNF-α Inhibitors in Therapeutics. Int J Mol Sci. 2021 Mar

2. Wang Y et al. The role of IL-1β and TNF-α in intervertebral disc degeneration. Biomed Pharmacother. 2020 Nov

Subcellular Location

Cell membrane; Secreted.

UNIPROT

SwissProt: P16599 Rat

Synonyms

APC1 antibody

APC1 protein antibody

Cachectin antibody

DIF antibody

Differentiation inducing factor antibody

Macrophage cytotoxic factor antibody

Tnf antibody

TNF superfamily member 2 antibody

TNF superfamily, member 2 antibody

TNF, macrophage derived antibody

Expand

Images

  • Sandwich ELISA analysis of Rat TNF-α matched pair antibodies<br /><br />Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (<a href="/products/HA722170" style="font-weight: bold;text-decoration: underline;">HA722170</a>) diluted in carbonate/bicarbonate buffer, at a concentration of 2 µg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1%BSA blocking buffer, and incubated with serial diluted Recombinant Rat TNF-α protein (<a href="/products/HA210538" style="font-weight: bold;text-decoration: underline;">HA210538</a>) starting from 1000 pg/ml to 0 pg/ml and detect antibody (<a href="/products/HA722171" style="font-weight: bold;text-decoration: underline;">HA722171</a>, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

    Sandwich ELISA analysis of Rat TNF-α matched pair antibodies

    Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722170) diluted in carbonate/bicarbonate buffer, at a concentration of 2 µg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1%BSA blocking buffer, and incubated with serial diluted Recombinant Rat TNF-α protein (HA210538) starting from 1000 pg/ml to 0 pg/ml and detect antibody (HA722171, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

  • Conditioned media was harvested after 3 days. TNF-α was measured in 100% unstimulated and LPS stimulated rat spleen cell supernatant. The concentrations of TNF-α were interpolated from the TNF-α standard curves. The mean TNF-α concentration was determined to be 124pg/mL in LPS stimulated rat spleen cell supernatant. There was no detectable signal in unstimulated supernatant.

    Conditioned media was harvested after 3 days. TNF-α was measured in 100% unstimulated and LPS stimulated rat spleen cell supernatant. The concentrations of TNF-α were interpolated from the TNF-α standard curves. The mean TNF-α concentration was determined to be 124pg/mL in LPS stimulated rat spleen cell supernatant. There was no detectable signal in unstimulated supernatant.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"