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Anti-TNF alpha antibody [EPR19147] ab183218 is a rabbit monoclonal antibody that is used in TNF alpha western blotting and immunofluorescence. Suitable for human and mouse samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with TNF knockout cell line validation
- Antibody clone EPR19147 is cited in over 180 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.

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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

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1 product for Alternative Version

Target data

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. Impairs regulatory T-cells (Treg) function in individuals with rheumatoid arthritis via FOXP3 dephosphorylation. Upregulates the expression of protein phosphatase 1 (PP1), which dephosphorylates the key 'Ser-418' residue of FOXP3, thereby inactivating FOXP3 and rendering Treg cells functionally defective (PubMed:23396208). Key mediator of cell death in the anticancer action of BCG-stimulated neutrophils in combination with DIABLO/SMAC mimetic in the RT4v6 bladder cancer cell line (PubMed:22517918, PubMed:16829952, PubMed:23396208). Induces insulin resistance in adipocytes via inhibition of insulin-induced IRS1 tyrosine phosphorylation and insulin-induced glucose uptake. Induces GKAP42 protein degradation in adipocytes which is partially responsible for TNF-induced insulin resistance (By similarity). Plays a role in angiogenesis by inducing VEGF production synergistically with IL1B and IL6 (PubMed:12794819).The TNF intracellular domain (ICD) form induces IL12 production in dendritic cells.

Alternative names

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

Recommended products

Anti-TNF alpha antibody [EPR19147] ab183218 is a rabbit monoclonal antibody that is used in TNF alpha western blotting and immunofluorescence. Suitable for human and mouse samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with TNF knockout cell line validation
- Antibody clone EPR19147 is cited in over 180 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

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

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)

    Immunocytochemistry/ Immunofluorescence analysis of RAW264.7(Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml LPS for 7 h and 1?g/ml BFA for the last 3h cells labeling TNF alpha with purified ab183218 at 1/5000 dilution?0.09 ©:g/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 ©:g/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 ©:g/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 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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