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Anti-TNF alpha antibody [EPR19147] (ab183218) is a rabbit monoclonal antibody detecting TNF alpha in Western Blot, IP, ICC/IF, ELISA. Suitable for Human, Mouse.



- KO validated for confirmed specificity

- Biophysical QC for unrivalled batch-batch consistency

- Over 100 publications


Images

Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [EPR19147] (AB183218), expandable thumbnail
  • ELISA - Anti-TNF alpha antibody [EPR19147] (AB183218), expandable thumbnail
  • Western blot - Anti-TNF alpha antibody [EPR19147] (AB183218), expandable thumbnail
  • Western blot - Anti-TNF alpha antibody [EPR19147] (AB183218), expandable thumbnail
  • Western blot - Anti-TNF alpha antibody [EPR19147] (AB183218), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Abcam Recommends

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPELISAWBICC/IF
Human
Tested
Expected
Tested
Expected
Mouse
Expected
Expected
Tested
Tested
Synthetic peptide - Human
Not recommended
Tested
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
1/40
Notes

-

Expected
Expected

Species
Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Expected
Expected

Species
Human, Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/1000
Notes

1/1000 dilution: Detects a band of approximately 26 kda (predicted molecular weight: 25 kDa).

Species
Human
Dilution info
1/1000 - 1/2000
Notes

1/1000 dilution: Detects a band of approximately 26 kda (predicted molecular weight: 25 kDa).

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/5000
Notes

-

Expected
Expected

Species
Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

1 product for Alternative Version

Target data

Function

The TNF protein, primarily secreted by macrophages, binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It induces cell death in specific tumor cell lines and acts as a potent pyrogen, causing fever directly or by stimulating interleukin-1 secretion. TNF is implicated in cachexia induction and can stimulate cell proliferation and differentiation under certain conditions. It impairs regulatory T-cells (Treg) function in rheumatoid arthritis patients through FOXP3 dephosphorylation, upregulating protein phosphatase 1 (PP1), which dephosphorylates 'Ser-418' of FOXP3, inactivating FOXP3 and leading to defective Treg cells. TNF is a key mediator of cell death in the anticancer effect of BCG-stimulated neutrophils with DIABLO/SMAC mimetic in the RT4v6 bladder cancer cell line. It induces insulin resistance in adipocytes by inhibiting insulin-induced IRS1 tyrosine phosphorylation and glucose uptake. TNF plays a role in angiogenesis by inducing VEGF production with IL1B and IL6. Additionally, the TNF intracellular domain (ICD) form stimulates IL12 production in dendritic cells. This supplementary information is collated from multiple sources and compiled automatically.

Alternative names

TNFA, TNFSF2, TNF, Tumor necrosis factor, Cachectin, TNF-alpha, Tumor necrosis factor ligand superfamily member 2, TNF-a

Recommended products

Anti-TNF alpha antibody [EPR19147] (ab183218) is a rabbit monoclonal antibody detecting TNF alpha in Western Blot, IP, ICC/IF, ELISA. Suitable for Human, Mouse.



- KO validated for confirmed specificity

- Biophysical QC for unrivalled batch-batch consistency

- Over 100 publications

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR19147
Purification technique
Affinity purification Protein A
Specificity

We recommend ab183218 to detect TNF alpha in Western blot, as it is more sensitive than ab215188. Stimulation is required for the detection of TNF alpha in most samples.

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Product Specifications
Anti-TNF alpha antibody [EPR19147] (ab183218) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in ELISA, ICC/IF, IP, WB in human, mouse samples.
Anti-TNF alpha antibody [EPR19147] (ab183218) specifically detects TNF alpha (UniProt ID: P01375; Molecular weight: 26kDa) and is sold in 100 µL and 1 mL selling sizes.

Quality and Validation
Abcam's high quality manufacturing and validation processes ensure Anti-TNF alpha antibody [EPR19147] (ab183218) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
The specificity of Anti-TNF alpha antibody [EPR19147] (ab183218) has been confirmed by testing in knockout samples.
Anti-TNF alpha antibody [EPR19147] (ab183218) has been cited over 104 times in peer reviewed journals and is trusted by the scientific community.


Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

6 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [EPR19147] (ab183218), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [EPR19147] (ab183218)

    TNF alpha Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-TNF alpha antibody

    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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (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.

  • ELISA - Anti-TNF alpha antibody [EPR19147] (ab183218), expandable thumbnail

    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), expandable thumbnail

    Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218)

    Blocking/Dilution buffer : 5% NFDM/TBST.

    All lanes: Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218) at 1/2000 dilution

    Lane 1: TPA pretreated THP-1 (Human monocytic leukemia cell line) whole cell lysate at 20 µg

    Lane 2: TPA pretreated THP-1 (Human monocytic leukemia cell line) whole cell lysate treated with 100 ng/ml LPS for 7 hours at 20 µg

    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 at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 25 kDa

    Observed band size: 26 kDa

    Exposure time: 3min

  • Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218), expandable thumbnail

    Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218)

    Lanes 1 - 6: Merged signal (red and green). Green - ab183218 observed at 26 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - 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 Anti-alpha Tubulin antibody [DM1A] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

    All lanes: Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218) at 1/1000 dilution

    Lane 1: Wild-type THP-1 Brefeldin A (Brefeldin A, Inhibitor of ADP-ribosylation factor ab120299) treated (5 µg/ml, 4 h) cell lysate at 30 µg

    Lane 2: Wild-type THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (Brefeldin A, Inhibitor of ADP-ribosylation factor ab120299) treated (5 µg/ml, 4 h) cell lysate at 30 µg

    Lane 2: Western blot - Human TNF knockout THP-1 cell line (Human TNF knockout THP-1 cell line ab273761)

    Lane 3: TNF alpha knockout THP-1 Brefeldin A (Brefeldin A, Inhibitor of ADP-ribosylation factor ab120299) treated (5 µg/ml, 4 h) cell lysate at 30 µg

    Lane 4: TNF alpha knockout THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (Brefeldin A, Inhibitor of ADP-ribosylation factor ab120299) treated (5 µg/ml, 4 h) cell lysate at 30 µg

    Lane 5: U937 PMA treated (10 mM, 2 days) plus 16 h no treatment and Brefeldin A (Brefeldin A, Inhibitor of ADP-ribosylation factor ab120299) treated (5 µg/ml, 4 h) cell lysate at 30 µg

    Lane 6: U937 PMA treated (10 mM, 2 days) and LPS treated (1 µg/ml, 16 h) plus Brefeldin A (Brefeldin A, Inhibitor of ADP-ribosylation factor ab120299) treated (5 µg/ml, 4 h) cell lysate at 30 µg

    Performed under reducing conditions.

    Predicted band size: 25 kDa

    Observed band size: 26 kDa

  • Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218), expandable thumbnail

    Western blot - Anti-TNF alpha antibody [EPR19147] (ab183218)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The 33kDa band probably is the dimeric intermediate form that has been described in the literature (PMID: 9933416).

    All lanes: Western blot - 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 at 20 µg

    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 at 20 µg

    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 at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 25 kDa

    Observed band size: 26 kDa, 33 kDa

    Exposure time: 3s

  • Immunoprecipitation - Anti-TNF alpha antibody [EPR19147] (ab183218), expandable thumbnail

    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) (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 (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.

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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