Pig (Porcine) Tumor Necrosis Factor ELISA Kit

Recommended Tumor Necrosis Factor Product (supplied by: Log in to see )

Antigen
Tumor Necrosis Factor (TNF) ELISA Kits
  • TNF
  • tnf
  • dif
  • tnfa
  • xtnf
  • TNF-a
  • tnfsf2
  • tnf-alpha
  • TNF-alpha
  • DIF
  • TNFA
  • TNFSF2
  • TNFalpha
  • Tnfa
  • Tnfsf1a
  • RATTNF
  • TNFa
  • cTNF
  • TNF-ALPHA
  • Cachectin
  • Tnf-alpha
  • tnfa-like
  • tumor necrosis factor (TNF superfamily, member 2)
  • tumor necrosis factor
  • tumour necrosis factor alpha-like
  • tumor necrosis factor alpha
  • lipopolysaccharide-induced TNF factor
  • tumor necrosis factor b (TNF superfamily, member 2)
  • TNF
  • tnf
  • tnf-alpha
  • LITAF
  • Tnf
  • tnfb
Reactivity
Pig (Porcine)
Alternatives
Kits with alternative reactivity to:
85
66
52
19
14
12
12
10
10
10
9
7
4
3
3
2
1
1
Method Type
Sandwich ELISA
Detection Range
20 pg/mL-10 ng/mL
Minimum Detection Limit
20 pg/mL
Application
ELISA
Options
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Catalog No. ABIN625441
$ 476.30
Plus shipping costs $45.00
Relevance Score ABIN Method Type Sample Type Detection Range Detection Minimum Supplier References Details
10.215476 ABIN415858 Sandwich ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 15.6-1000 pg/mL 15.6 pg/mL Log in to see 3
10.215476 ABIN858549 Sandwich ELISA Serum, Plasma, Cell Culture Supernatant, Tissue Homogenate 0.047-3 ng/mL 0.047 ng/mL Log in to see
10.215476 ABIN456702 Sandwich ELISA Cell Culture Supernatant, Serum, Tissue Homogenate, Plasma 31.2-2000 pg/mL 31.2 pg/mL Log in to see
1 ABIN3032860 Competition ELISA Cell Culture Supernatant, Plasma, Serum 13.7-10000 pg/mL 13.7 pg/mL Log in to see
1 ABIN1117652 Sandwich ELISA Serum, Plasma, Biological Fluids 31.25-2000 pg/mL 31.25 pg/mL Log in to see
1 ABIN1572788 Sandwich ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 62.5-4000 pg/mL 62.5 pg/mL Log in to see
1 ABIN779971 Competition ELISA Serum, Plasma, Cell Culture Supernatant, Tissue Homogenate, Body Fluids 50-1000 pg/mL 50 pg/mL Log in to see
1 ABIN5522443 Sandwich ELISA Plasma, Serum, Biological Fluids, Tissue Homogenate 31.25-2000 pg/mL 31.25 pg/mL Log in to see
1 ABIN955998 Sandwich ELISA 15.6-1000 pg/mL 15.6 pg/mL Log in to see
1 ABIN4987126 Sandwich ELISA Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (citrate), Plasma (EDTA) 0.03-2.0 ng/mL 0.03 ng/mL Log in to see
1 ABIN2648840 Sandwich ELISA Serum, Plasma, Cell Culture Cells Log in to see
1 ABIN2748911 Sandwich ELISA Cell Culture Supernatant, Serum, Plasma, Tissue Homogenate 31.3-2000 pg/mL 31.3 pg/mL Log in to see
1 ABIN4987145 Sandwich ELISA Cell Culture Supernatant, Plasma, Tissue Homogenate, Serum 0.047-3 ng/mL 0.047 ng/mL Log in to see
1 ABIN4973389 Sandwich ELISA Cell Culture Supernatant, Plasma, Tissue Homogenate, Serum, Cell Lysate, Biological Fluids 62.5-4000 pg/mL 62.5 pg/mL Log in to see
1 ABIN2540085 Sandwich ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 15.63-1000 pg/mL 15.63 pg/mL Log in to see
1 ABIN2685694 Sandwich ELISA Cell Culture Supernatant, Plasma, Serum Log in to see
1 ABIN2952130 31.25-2000 pg/mL 31.25 pg/mL Log in to see
1 ABIN2109940 Competition ELISA Log in to see
1 ABIN5072432 Sandwich ELISA Plasma, Serum, Tissue Homogenate, Biological Fluids Log in to see

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General

Antigen Tumor Necrosis Factor (TNF) ELISA Kits
Reactivity Pig (Porcine)
Kits with alternative reactivity to:
(85), (66), (52), (19), (14), (12), (12), (10), (10), (10), (9), (7), (4), (3), (3), (2), (1), (1)
Method Type Sandwich ELISA
Detection Range 20 pg/mL-10 ng/mL
Minimum Detection Limit 20 pg/mL
Application ELISA
Pubmed 6 references available
Supplier Log in to see

Product Details Tumor Necrosis Factor ELISA Kit

Target details Application Details Handling ProductDetails: References for Tumor Necrosis Factor Kit (ABIN625441) Images
Purpose Porcine TNF alpha ELISA Kit for cell culture supernatants, plasma, and serum samples.
Sample Type Plasma, Cell Culture Supernatant, Serum
Detection Method Colorimetric
Analytical Method Quantitative
Specificity The sandwich ELISA antibody pair detects Porcine TNF-alpha.
Sensitivity 20 pg/mL
Characteristics
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data
Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Material not included
  • Distilled or deionized water
  • Precision pipettes to deliver 2 μL to 1 μL volumes
  • Adjustable 1-25 μL pipettes for reagent preparation
  • 100 μL and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis

Target details

Product Details Tumor Necrosis Factor ELISA Kit Application Details Handling ProductDetails: References for Tumor Necrosis Factor Kit (ABIN625441) Images back to top
Antigen
Alternative Name TNF-alpha (TNF ELISA Kit Abstract)
Background Tumor necrosis factor (Cachectin) (TNF-alpha) (Tumor necrosis factor ligand superfamily member 2) (TNF-a)
Gene ID 397086
UniProt P23563
Research Area Virology, Atherosclerosis, Immunology, Receptors, Cytokines, Growth Factors, Innate Immunity, Cancer, Cardiovascular, Inflammation
Pathways NF-kappaB Signaling, Apoptosis, Caspase Cascade in Apoptosis, Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Positive Regulation of Endopeptidase Activity, Hepatitis C, Protein targeting to Nucleus

Application Details

Product Details Tumor Necrosis Factor ELISA Kit Target details Handling ProductDetails: References for Tumor Necrosis Factor Kit (ABIN625441) Images back to top
Application Notes Recommended Dilution for serum and plasma samples2 fold
Sample Volume 100 μL
Plate Pre-coated
Protocol
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 μL of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4 °C.
  4. Add 100 μL of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 μL of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 μL of Stop Solution to each well.
  11. Read at 450 nm immediately.
Reagent Preparation 1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
2. Sample dilution: If your samples need to be diluted, Assay Diluent C (Item L) should be used for dilution of serum/plasma/culture supernatants. Suggested dilution for normal serum/plasma: 2 fold*. * Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
3. Assay Diluent B (Item E) should be diluted 5-fold with deionized or distilled water before use.
4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µL Assay Diluent C (Item L) into Item C vial to prepare a 40 ng/mL standard solution. Dissolve the powder thoroughly by a gentle mix. Add 200 µL TNF-alpha standard from the vial of Item C, into a tube with 600 µL Assay Diluent C to prepare a 10 ng/mL standard solution. Pipette 400myl Assay Diluent C into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent C serves as the zero standard (0 ng/mL). 200 µL 200 µL 200 µL 200 µL 200 µL 200myl 200 µL standard + 600 µL 10 3.333 1.111 0.370 0.123 0.041 0.014 0 ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL
5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B (Item E) into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 600-fold with 1x Assay Diluent B (Item E). For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 20 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent B to prepare a 600-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
Assay Procedure 1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
5. Discard the solution. Repeat the wash as in step
6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
7. Discard the solution. Repeat the wash as in step
8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
Calculation of Results Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent C Porcine TNF-alpha concentration (ng/mL) 0.01 0.1 1 10 O D =4 50 n m 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of TNF-alpha is typically less than 20 pg/mL.
Recovery: Recovery was determined by spiking TNF-alpha into porcine serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 111.7 102-121 Plasma 97.02 70-126 Cell culture media 132.1 123-142
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 89.04 94.57 103.3 Range ( %) 78-97 85-102 92-118 1:4 Average % of Expected 75.78 74.79 76.55 Range ( %) 67-85 67-86 68-88
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %
Assay Precision Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
Restrictions For Research Use only

Handling

Product Details Tumor Necrosis Factor ELISA Kit Target details Application Details ProductDetails: References for Tumor Necrosis Factor Kit (ABIN625441) Images back to top
Handling Advice Avoid repeated freeze-thaw cycles.
Storage -20 °C
Storage Comment The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Expiry Date 6 months

ProductDetails: References for Tumor Necrosis Factor Kit (ABIN625441)

Product Details Tumor Necrosis Factor ELISA Kit Target details Application Details Handling Images back to top
Product cited in:

Zong, Song, Wang, Xia, Hu, Han, Wang: "LFP-20, a porcine lactoferrin peptide, ameliorates LPS-induced inflammation via the MyD88/NF-κB and MyD88/MAPK signaling pathways." in: Developmental and comparative immunology, Vol. 52, Issue 2, pp. 123-31, 2015

Zhang, Hou, Zhong, Li, Chen, Li, Wen, Wang, Liu, Zhong: "Porcine reproductive and respiratory syndrome virus activates inflammasomes of porcine alveolar macrophages via its small envelope protein E." in: Virology, Vol. 442, Issue 2, pp. 156-62, 2013

Zhang, Hou, Zhong, Li, Chen, Li, Wen, Wang, Liu, Zhong: "Porcine reproductive and respiratory syndrome virus activates inflammasomes of porcine alveolar macrophages via its small envelope protein E." in: Virology, Vol. 442, Issue 2, pp. 156-62, 2013

Zhang, Hou, Zhong, Li, Chen, Li, Wen, Wang, Liu, Zhong: "Porcine reproductive and respiratory syndrome virus activates inflammasomes of porcine alveolar macrophages via its small envelope protein E." in: Virology, Vol. 442, Issue 2, pp. 156-62, 2013

Zhang, Hou, Zhong, Li, Chen, Li, Wen, Wang, Liu, Zhong: "Porcine reproductive and respiratory syndrome virus activates inflammasomes of porcine alveolar macrophages via its small envelope protein E." in: Virology, Vol. 442, Issue 2, pp. 156-62, 2013

Galindo, Ayllón, Smrdel, Boadella, Beltrán-Beck, Mazariegos, García, de la Lastra, Avsic-Zupanc, Kocan, Gortazar, de la Fuente: "Gene expression profile suggests that pigs (Sus scrofa) are susceptible to Anaplasma phagocytophilum but control infection." in: Parasites & vectors, Vol. 5, pp. 181, 2012

Images

Product Details Tumor Necrosis Factor ELISA Kit Target details Application Details Handling ProductDetails: References for Tumor Necrosis Factor Kit (ABIN625441) back to top
Supplier Images
ELISA image for Tumor Necrosis Factor ELISA Kit (TNF) (ABIN625441) Tumor Necrosis Factor (TNF) ELISA Kit