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Rabbit Recombinant Monoclonal TNF alpha antibody. Suitable for IP, WB, Flow Cyt (Intra) and reacts with Human samples. Cited in 8 publications.

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Images

Immunoprecipitation - Anti-TNF alpha antibody [EPR22598-212] (AB255275), expandable thumbnail
  • Western blot - Anti-TNF alpha antibody [EPR22598-212] (AB255275), expandable thumbnail
  • Western blot - Anti-TNF alpha antibody [EPR22598-212] (AB255275), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [EPR22598-212] (AB255275), 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
IPWBIHC-PICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Not recommended
Not recommended
Tested

Tested
Tested

Species
Human
Dilution info
1/30
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/700
Notes

-

Associated Products

Select an associated product type

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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

Rabbit Recombinant Monoclonal TNF alpha antibody. Suitable for IP, WB, Flow Cyt (Intra) and reacts with Human samples. Cited in 8 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR22598-212
Purification technique
Affinity purification Protein A
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

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.

4 product images

  • Immunoprecipitation - Anti-TNF alpha antibody [EPR22598-212] (ab255275), expandable thumbnail

    Immunoprecipitation - Anti-TNF alpha antibody [EPR22598-212] (ab255275)

    TNF alpha was immunoprecipitated from 0.35 mg of THP-1 (human monocytic leukemia cell line) (treated with 80nM TPA overnight, replaced the culture medium with 100 ng/ml Lipopolysaccharides (LPS) for 6 hours with addition of 300ng/ml Brefeldin A (BFA) for the last 3 hours) whole cell lysate with ab255275 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab255275 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used as secondary antibody at 1/5000 dilution.

    Lane 1: THP-1 (treated as above) whole cell lysate 10 μg (Input).
    Lane 2: ab255275 IP in THP-1 (treated as avove) whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab255275 in THP-1 (treated as above) whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 15 seconds.

    All lanes: Immunoprecipitation - Anti-TNF alpha antibody [EPR22598-212] (ab255275)

    Predicted band size: 25 kDa

    Observed band size: 25 kDa

  • Western blot - Anti-TNF alpha antibody [EPR22598-212] (ab255275), expandable thumbnail

    Western blot - Anti-TNF alpha antibody [EPR22598-212] (ab255275)

    Lanes 1 - 6: Merged signal (red and green). Green - ab255275 observed at 26 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab255275 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 ab255275 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 [EPR22598-212] (ab255275) 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 [EPR22598-212] (ab255275), expandable thumbnail

    Western blot - Anti-TNF alpha antibody [EPR22598-212] (ab255275)

    Blocking and dilution buffer: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature (PMID: 9657756).

    All lanes: Western blot - Anti-TNF alpha antibody [EPR22598-212] (ab255275) at 1/1000 dilution

    Lane 1: THP-1 (human monocytic leukemia cell line) (treated with 80nM 12-O-Tetradecanoylphorbol-13-acetate (TPA) overnight) whole cell lysate at 20 µg

    Lane 2: THP-1 (treated with 80nM TPA overnight, replaced the culture medium with 100 ng/ml lipopolysaccharides (LPS) for 6 hours with addition of 300ng/ml Brefeldin A (BFA) for the last 3 hours) whole cell lysate 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: 25 kDa

    Exposure time: 3min

  • Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [EPR22598-212] (ab255275), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [EPR22598-212] (ab255275)

    Intracellular flow cytometric analysis of2% paraformaldehyde-fixed, 0.1% Tween 20 permeabilized Human peripheral blood mononuclear cell (PBMC) treated with 50ng/ml PMA, 250ng/ml ionomycin and 300ng/ml Brefeldin A for 16h, labeling TNF alpha with ab255275 at 1/700 dilution (Right) compared with Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Left).

    Goat Anti-Rabbit IgG Fc (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG Fc (Alexa Fluor® 488) preadsorbed ab150097), at 1/5000 dilution was used as the secondary antibody.

    Cells were surface stained with anti-CD3 conjugated to Alexa Fluor® 647. Then fixed with 2% PFA for 10min and intracellularly stained with rabbit IgG (Left) or ab236836 (Right).

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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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