Mouse/Rat TNF alpha Recombinant Rabbit Monoclonal Antibody [PSH03-30]
cat.: HA721975
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Mouse, Rat |
| Applications: | ELISA(Det) |
| Clonality: | Monoclonal |
| Clone number: | PSH03-30 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4). |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 25.8 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within Rat TNF alpha aa 80-235 (P16599). |
| Positive control: | Recombinant rat TNF alpha protein, Recombinant mouse TNF alpha protein. |
| Subcellular location: | Secreted. |
| Recommended Dilutions:
ELISA(Det) |
Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH03-29] to Mouse/Rat TNF alpha (Capture) (HA721974) and recombinant standard Mouse TNF alpha (HA210527) and Rat TNF alpha (HA210538) as the standard. The reference range value is 12.3-3000 pg/ml. |
| Uniprot #: | SwissProt: P06804 Mouse | P16599 Rat |
| Alternative names: | APC1 APC1 protein Cachectin DIF Differentiation inducing factor Macrophage cytotoxic factor Tnf TNF superfamily member 2 TNF superfamily, member 2 TNF, macrophage derived TNF, monocyte derived TNF-a TNF-alpha TNFA TNFA_HUMAN TNFSF2 Tumor necrosis factor Tumor necrosis factor (TNF superfamily member 2) Tumor necrosis factor alpha Tumor necrosis factor ligand superfamily member 2 Tumor Necrosis Factor, Membrane Form Tumor necrosis factor, soluble form |
Images
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Fig1:
Sandwich ELISA analysis of rat TNF alpha matched pair antibodies Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA721974) 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 rat TNF alpha protein (HA210538) starting from 3000 pg/ml to 0 pg/ml and detect antibody (HA721975) -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. |
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Fig2:
Sandwich ELISA analysis of mouse TNF alpha matched pair antibodies Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA721974) 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 mouse TNF alpha protein (HA210527) starting from 3000 pg/ml to 0 pg/ml and detect antibody (HA721975)-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. |
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Fig3: Conditioned media was harvested after 72 hours. 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 128 pg/mL in LPS stimulated rat spleen cell supernatant. There was no detectable signal in unstimulated supernatant. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.