Rabbit Monoclonal TNF alpha antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Mouse, Human samples. Cited in 43 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IP | WB | ICC/IF | Flow Cyt (Intra) | |
---|---|---|---|---|
Human | Expected | Tested | Expected | Expected |
Mouse | Tested | Tested | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/30 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
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 |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
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. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Select an associated product type
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.
TNFA, TNFSF2, TNF, Tumor necrosis factor, Cachectin, TNF-alpha, Tumor necrosis factor ligand superfamily member 2, TNF-a
Rabbit Monoclonal TNF alpha antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Mouse, Human samples. Cited in 43 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR20972
Affinity purification Protein A
The protein level of TNF alpha in normal samples is very weak. The TNF alpha expression must be stimulated.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our 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.
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
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
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
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.
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
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.
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
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
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com