Recombinant Anti-TNF alpha antibody [EPR19147] (ab183218)
![Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218)](https://www.abcam.com/ps/products/183/ab183218/Images/ab183218-450485-anti-tnf-alpha-antibody-epr19147-western-blot-wildtype-thp1-brefeldin-a-treated-5-gml-4-h-wildtype-thp1-lps-treate.jpg "Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19147] to TNF alpha
- Suitable for: WB, IP, ELISA, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Overview
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Product name
Anti-TNF alpha antibody [EPR19147]
See all TNF alpha primary antibodies -
Description
Rabbit monoclonal [EPR19147] to TNF alpha -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF MouseIP HumanWB MouseHuman
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: TPA pretreated THP-1 treated with 100 ng/ml LPS for 7 hours and TPA pretreated THP-1 treated with 100 ng/ml LPS for 4 hours, then added 1 µg/ml BFA for 3 hours whole cell lysates; TPA pretreated RAW 264.7, TPA pretreated RAW 264.7 treated with 100 ng/ml LPS for 7 hours and TPA pretreated RAW 264.7 treated with 100 ng/ml LPS for 4 hours, then added 1 µg/ml BFA for 3 hours whole cell lysates. IP: TPA pretreated THP-1 treated with 100 ng/ml LPS for 4 hours, then added 1 µg/ml BFA for 3 hours cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19147 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab183218 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| ICC/IF |
Mouse
|
| IP |
Human
|
| WB |
Mouse
Human
|
| Application | Abreviews | Notes |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 26, 14 kDa (predicted molecular weight: 25 kDa).
|
|
| IP |
1/40.
|
|
| ELISA |
Use at an assay dependent concentration.
|
|
| ICC/IF |
1/5000.
|
| Notes |
|---|
|
WB
1/1000. Detects a band of approximately 26, 14 kDa (predicted molecular weight: 25 kDa). |
|
IP
1/40. |
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ELISA
Use at an assay dependent concentration. |
|
ICC/IF
1/5000. |
Target
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Function
Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation. -
Involvement in disease
Genetic variations in TNF are a cause of susceptibility psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis). -
Sequence similarities
Belongs to the tumor necrosis factor family. -
Post-translational
modificationsThe soluble form derives from the membrane form by proteolytic processing.
The membrane form, but not the soluble form, is phosphorylated on serine residues. Dephosphorylation of the membrane form occurs by binding to soluble TNFRSF1A/TNFR1.
O-glycosylated; glycans contain galactose, N-acetylgalactosamine and N-acetylneuraminic acid. -
Cellular localization
Secreted and Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 7124 Human
- Entrez Gene: 21926 Mouse
- Omim: 191160 Human
- SwissProt: P01375 Human
- SwissProt: P06804 Mouse
- Unigene: 241570 Human
- Unigene: 1293 Mouse
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Alternative names
- APC1 antibody
- APC1 protein antibody
- Cachectin antibody
see all
Images
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Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218)All lanes : Anti-TNF alpha antibody [EPR19147] (ab183218) at 1/1000 dilution
Lane 1 : Wild-type THP-1 Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
Lane 2 : Wild-type THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
Lane 3 : TNF alpha knockout THP-1 Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
Lane 4 : TNF alpha knockout THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
Lane 5 : U937 PMA treated (10 mM, 2 days) plus 16 h no treatment and Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
Lane 6 : U937 PMA treated (10 mM, 2 days) and LPS treated (1 µg/ml, 16 h) plus Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
Lysates/proteins at 30 µg per lane.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?Lanes 1 - 6: Merged signal (red and green). Green - ab183218 observed at 26 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab183218 was shown to react with TNF alpha in THP-1 wild-type cells in Western blot with loss of signal observed in TNF knockout sample. Wild-type and TNF knockout THP-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab183218 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [EPR19147] (ab183218)](https://www.abcam.com/ps/products/183/ab183218/Images/ab183218-330915-anti-tnf-alpha-antibody-epr19147-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [EPR19147] (ab183218)Immunocytochemistry/ Immunofluorescence analysis of RAW264.7(Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml LPS for 7 h and 1µg/ml BFA for the last 3h cells labeling TNF alpha with purified ab183218 at 1/5000 dilution (0.09 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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![ELISA - Anti-TNF alpha antibody [EPR19147] (ab183218)](https://www.abcam.com/ps/products/183/ab183218/Images/ab183218-330894-anti-tnf-alpha-antibody-epr19147-elisa.jpg)
ELISA - Anti-TNF alpha antibody [EPR19147] (ab183218)ELISA using ab183218 between 0.1 and 1000 ng/ml to detect TNF alpha peptide. Secondary used was Alkaline Phosphatase AffiniPure Goat Anti-Rabbit IgG (H+L) 1/2000. -
![Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218)](https://www.abcam.com/ps/products/183/ab183218/Images/ab183218-266031-anti-tnf-alpha-antibody-epr19147-western-blot.jpg)
Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218)All lanes : Anti-TNF alpha antibody [EPR19147] (ab183218) at 1/2000 dilution
Lane 1 : TPA pretreated THP-1 (Human monocytic leukemia cell line) whole cell lysate
Lane 2 : TPA pretreated THP-1 (Human monocytic leukemia cell line) whole cell lysate treated with 100 ng/ml LPS for 7 hours
Lane 3 : TPA pretreated THP-1 (Human monocytic leukemia cell line) whole cell lysate treated with 100 ng/ml LPS for 4 hours, then added 1 µg/ml BFA for 3 hours
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 25 kDa
Observed band size: 14,26 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The 14kDa band probably is the degradable inactive fragment that has been decribed by literature (PMID: 9933416).
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![Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218)](https://www.abcam.com/ps/products/183/ab183218/Images/ab183218-266030-anti-tnf-alpha-antibody-epr19147-western-blot.jpg)
Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218)All lanes : Anti-TNF alpha antibody [EPR19147] (ab183218) at 1/1000 dilution
Lane 1 : TPA pretreated RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 2 : TPA pretreated RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate treated with 100 ng/ml LPS for 7 hours
Lane 3 : TPA pretreated RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate treated with 100 ng/ml LPS for 4 hours, then added 1 µg/ml BFA for 3 hours
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 25 kDa
Observed band size: 14,26,33 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The 33kDa band probably is the dimeric intermediate form that has been described in the literature (PMID: 9933416).
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![Immunoprecipitation - Anti-TNF alpha antibody [EPR19147] (ab183218)](https://www.abcam.com/ps/products/183/ab183218/Images/ab183218-266029-anti-tnf-alpha-antibody-epr19147-immunoprecipitation.jpg)
Immunoprecipitation - Anti-TNF alpha antibody [EPR19147] (ab183218)TNF alpha was immunoprecipitated from 0.35 mg of TPA pretreated THP-1 (Human monocytic leukemia cell line) whole cell lysate treated with 100 ng/ml LPS for 4 hours, then added 1 μg/ml BFA for 3 hours with ab183218 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab183218 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: TPA pretreated THP-1 whole cell lysate treated with 100 ng/ml LPS for 4 hours, then added 1 μg/ml BFA for 3 hours 10µg (Input).
Lane 2: ab183218 IP in TPA pretreated THP-1 whole cell lysate treated with 100 ng/ml LPS for 4 hours, then added 1 μg/ml BFA for 3 hours.
Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab183218 in TPA pretreated THP-1 whole cell lysate treated with 100 ng/ml LPS for 4 hours, then added 1 μg/ml BFA for 3 hours.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
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![Anti-TNF alpha antibody [EPR19147] (ab183218)](https://www.abcam.com/ps/products/183/ab183218/Images/ab183218-8-benefits-of-recombinant-antibodies.png)
Anti-TNF alpha antibody [EPR19147] (ab183218)
Protocols
Datasheets and documents
Certificate of Compliance
References (7)
ab183218 has been referenced in 7 publications.
- Jo ES et al. Sulfur Compounds Inhibit High Glucose-Induced Inflammation by Regulating NF-?B Signaling in Human Monocytes. Molecules 25:N/A (2020). PubMed: 32429534
- Bai YF et al. The flavonoid-rich Quzhou Fructus Aurantii extract modulates gut microbiota and prevents obesity in high-fat diet-fed mice. Nutr Diabetes 9:30 (2019). PubMed: 31645541
- Yambire KF et al. Impaired lysosomal acidification triggers iron deficiency and inflammation in vivo. Elife 8:N/A (2019). PubMed: 31793879
- Hong S et al. Sevoflurane induces apoptosis of isolated placental trophoblast cells and stimulates expressions of TNF-a and IL-6. Am J Transl Res 11:7084-7093 (2019). PubMed: 31814911
- Liang Z & Ren C Emodin attenuates apoptosis and inflammation induced by LPS through up-regulating lncRNA TUG1 in murine chondrogenic ATDC5 cells. Biomed Pharmacother 103:897-902 (2018). WB ; Mouse . PubMed: 29710506
- Yang QC et al. A murine model of dry eye induced by topical administration of erlotinib eye drops. Int J Mol Med 41:1427-1436 (2018). PubMed: 29286080
- Tan G et al. Air pollutant particulate matter 2.5 induces dry eye syndrome in mice. Sci Rep 8:17828 (2018). PubMed: 30546125
