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Rabbit Monoclonal TNF alpha antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Mouse, Human samples. Cited in 43 publications.


Images

Western blot - Anti-TNF alpha antibody [EPR20972] (AB215188), expandable thumbnail
  • Western blot - Anti-TNF alpha antibody [EPR20972] (AB215188), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [EPR20972] (AB215188), expandable thumbnail
  • Immunoprecipitation - Anti-TNF alpha antibody [EPR20972] (AB215188), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [EPR20972] (AB215188), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 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
IPWBICC/IFFlow Cyt (Intra)
Human
Expected
Tested
Expected
Expected
Mouse
Tested
Tested
Tested
Tested

Tested
Tested

Species

Mouse

Dilution info
1/30
Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info
1/1000
Notes

We recommend Anti-TNF alpha antibody [EPR19147] ab183218 to detect TNF alpha in Western blot, as it is more sensitive than ab215188

Species

Human

Dilution info
1/1000
Notes

We recommend Anti-TNF alpha antibody [EPR19147] ab183218 to detect TNF alpha in Western blot, as it is more sensitive than ab215188

Tested
Tested

Species

Mouse

Dilution info
1/100
Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info
1/600
Notes

Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

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. Up-regulates 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:16829952, PubMed:22517918, 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). Promotes osteoclastogenesis and therefore mediates bone resorption (By similarity).The TNF intracellular domain (ICD) form induces IL12 production in dendritic cells.

Alternative names

Recommended products

Rabbit Monoclonal TNF alpha antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Mouse, Human samples. Cited in 43 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

EPR20972

Purification technique

Affinity purification Protein A

Specificity

The protein level of TNF alpha in normal samples is very weak. The TNF alpha expression must be stimulated.

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

  • Western blot - Anti-TNF alpha antibody [EPR20972] (ab215188), expandable thumbnail

    Western blot - Anti-TNF alpha antibody [EPR20972] (ab215188)

    This Western blot image is a comparison between ab215188 and Anti-TNF alpha antibody [EPR19147] ab183218 tested under the same conditions. While ab215188 is suitable for WB for some samples, Anti-TNF alpha antibody [EPR19147] ab183218 was found to be more sensitive. False colour image of Western blot: Anti-TNF alpha antibody [EPR20972] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab215188 was shown to bind specifically to TNF alpha. A band was observed at 27 kDa in treated U937 cell lysates with no signal observed at this size without treatment. No signal was observed in wild-type THP-1 cell lysates or in TNF knockout cell line Human TNF knockout THP-1 cell line ab273761 (knockout cell lysate Human TNF knockout THP-1 cell lysate ab275507) with ab215188. However, a band was observed at 27 kDa in treated wild-type THP-1 cell lysates with Anti-TNF alpha antibody [EPR19147] ab183218. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were 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) at 1/20000 dilution.

    All lanes: Western blot - Anti-TNF alpha antibody [EPR20972] (ab215188) at 1/1000 dilution

    Lane 1: Wild-type THP-1 control: Brefeldin A (5 ug/mL, 4 h) cell lysate at 30 µg

    Lane 2: Wild-type treated THP-1: LPS (100 ng/mL, 16 h), Brefeldin A (5 ug/mL, last 4 h) cell lysate at 30 µg

    Lane 3: TNF alpha knockout THP-1 control: Brefeldin A (5 ug/mL, 4 h) cell lysate at 30 µg

    Lane 4: TNF alpha knockout THP-1 treated: LPS (100 ng/mL, 16 h), Brefeldin A (5 ug/mL, last 4 h) cell lysate at 30 µg

    Lane 5: U937 control: PMA (10 mM, 2 days), Brefeldin A (5 ug/mL, last 4 h) cell lysate at 30 µg

    Lane 6: U937 treated: PMA (10 mM, 2 days), LPS (1 ug/mL, last 16 h), Brefeldin A (5 ug/mL, last 4 h) cell lysate at 30 µg

    Performed under reducing conditions.

    Predicted band size: 25 kDa

    Observed band size: 27 kDa

  • Western blot - Anti-TNF alpha antibody [EPR20972] (ab215188), expandable thumbnail

    Western blot - Anti-TNF alpha antibody [EPR20972] (ab215188)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-TNF alpha antibody [EPR20972] (ab215188) at 1/1000 dilution

    Lane 1: Untreated RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus), whole cell lysate at 10 µg

    Lane 2: Untreated RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) treated with 100 ng/ml lipopolysaccharides (LPS) for 7 hours with addition of 1 μg/ml brefeldi at 10 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

    Developed using the ECL technique.

    Predicted band size: 25 kDa

    Observed band size: 17 kDa, 26 kDa, 33 kDa

    Exposure time: 15s

  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [EPR20972] (ab215188), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [EPR20972] (ab215188)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells, untreated or treated with 100 ng/ml lipopolysaccharides (LPS) for 7 hours with addition of 1 μg/ml brefeldin A (BFA) for the last 3 hours labeling TNF alpha with ab215188 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasmic staining was increased on RAW 264.7 cells when treated with 100 ng/ml lipopolysaccharides (LPS) for 7 hours with addition of 1 μg/ml brefeldin A (BFA) for the last 3 hours.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

  • Immunoprecipitation - Anti-TNF alpha antibody [EPR20972] (ab215188), expandable thumbnail

    Immunoprecipitation - Anti-TNF alpha antibody [EPR20972] (ab215188)

    TNF alpha was immunoprecipitated from 0.35 mg of RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) treated with 100 ng/ml lipopolysaccharides (LPS) for 7 hours with addition of 1 μg/ml brefeldin A (BFA) for the last 3 hours, whole cell lysate with ab215188 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab215188 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10,000 dilution

    Lane 1: RAW 264.7 treated with 100 ng/ml lipopolysaccharides (LPS) for 7 hours with addition of 1 μg/ml brefeldin A (BFA) for the last 3 hours, whole cell lysate 10 μg (Input).

    Lane 2: ab215188 IP in RAW 264.7 treated with 100 ng/ml lipopolysaccharides (LPS) for 7 hours with addition of 1 μg/ml brefeldin A (BFA) for the last 3 hours, whole cell lysate (+).

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab215188 in RAW 264.7 treated with 100 ng/ml lipopolysaccharides (LPS) for 7 hours with addition of 1 μg/ml brefeldin A (BFA) for the last 3 hours, whole cell lysate (-).

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

    All lanes: Immunoprecipitation - Anti-TNF alpha antibody [EPR20972] (ab215188)

    Predicted band size: 25 kDa

    Observed band size: 26 kDa, 33 kDa

  • Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [EPR20972] (ab215188), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [EPR20972] (ab215188)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized mouse splenocytes treated with 20ng/ml PMA, 1μg/ml Ionomycin and 10μM Brefeldin A for 6 hours labeling TNF alphawith ab215188 at 1/600 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

    Cells were surface stained with anti-mouse CD3, fixed with 4% PFA for 10 minutes, then permeabilized with 0.1% Tween-20 and intracellular stained with anti-rabbit IgG and ab215188. TNF alpha is mainly expressed in T cells (CD3+ population) while only a small population of CD3- cells can express TNF-alpha.

  • Western blot - Anti-TNF alpha antibody [EPR20972] (ab215188), expandable thumbnail

    Western blot - Anti-TNF alpha antibody [EPR20972] (ab215188)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-TNF alpha antibody [EPR20972] (ab215188) at 1/1000 dilution

    Lane 1: THP-1 (human monocytic leukemia cell line) differentiated with 100 nM TPA overnight, whole cell lysate at 10 µg

    Lane 2: THP-1 (human monocytic leukemia cell line) differentiated with 100 nM TPA overnight, then treated with 100 ng/ml lipopolysaccharides (LPS) for 7 hours with addition of 1 μg/ml brefeldin A (BFA) for the last 3 hours, whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

    Developed using the ECL technique.

    Predicted band size: 25 kDa

    Observed band size: 26 kDa

    Exposure time: 3min

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Product protocols

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