Rabbit Recombinant Monoclonal TNF alpha antibody. Carrier free. Suitable for IP, WB, Flow Cyt (Intra) and reacts with Human samples.
Images
![Immunoprecipitation - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (AB255288), expandable thumbnail](https://content.abcam.com/products/images/tnf-alpha-antibody-epr22598-212-bsa-and-azide-free-ab255288--immunoprecipitation-img147788.jpg/_jcr_content/renditions/webp-475.webp "Immunoprecipitation - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (AB255288)")
![Western blot - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (AB255288), expandable thumbnail](https://content.abcam.com/products/images/tnf-alpha-antibody-epr22598-212-bsa-and-azide-free-ab255288--western-blot-img47099.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (AB255288)")
![Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (AB255288), expandable thumbnail](https://content.abcam.com/products/images/tnf-alpha-antibody-epr22598-212-bsa-and-azide-free-ab255288--flow-cytometry-intracellular-img415544.jpg/_jcr_content/renditions/webp-475.webp "Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (AB255288)")
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
- Liquid
- Clonality
- Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Reactivity data
Select an application| IP | WB | IHC-P | ICC/IF | Flow Cyt (Intra) | |
|---|---|---|---|---|---|
| Human | Tested | Tested | Not recommended | Not recommended | Tested |
IP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
IHC-P
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
ICC/IF
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Flow Cyt (Intra)
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Associated Products
Select an associated product type
8 products for Alternative Product
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. Carrier free. Suitable for IP, WB, Flow Cyt (Intra) and reacts with Human samples.
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
- Liquid
- Clonality
- Monoclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Carrier free
- Yes
- Clone number
- EPR22598-212
- Purification technique
- Affinity purification Protein A
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- Do Not Freeze
Notes
ab255288 is the carrier-free version of Anti-TNF alpha antibody [EPR22598-212] ab255275.
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.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
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.
Not recommended
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.
3 product images
![Immunoprecipitation - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (ab255288), expandable thumbnail](https://content.abcam.com/products/images/tnf-alpha-antibody-epr22598-212-bsa-and-azide-free-ab255288--immunoprecipitation-img147788.jpg "Immunoprecipitation - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (ab255288)")
Immunoprecipitation - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (ab255288)
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 Anti-TNF alpha antibody [EPR22598-212] ab255275 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-TNF alpha antibody [EPR22598-212] 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: Anti-TNF alpha antibody [EPR22598-212] 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 Anti-TNF alpha antibody [EPR22598-212] ab255275 in THP-1 (treated as above) whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 15 seconds.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (Anti-TNF alpha antibody [EPR22598-212] ab255275).
All lanes: Immunoprecipitation - Anti-TNF alpha antibody [EPR22598-212] (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] - BSA and Azide free (ab255288), expandable thumbnail](https://content.abcam.com/products/images/tnf-alpha-antibody-epr22598-212-bsa-and-azide-free-ab255288--western-blot-img47099.jpg "Western blot - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (ab255288)")
Western blot - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (ab255288)
This data was developed using the same antibody clone in a different buffer formulation (Anti-TNF alpha antibody [EPR22598-212] ab255275).
Lanes 1 - 6: Merged signal (red and green). Green - Anti-TNF alpha antibody [EPR22598-212] 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.
Anti-TNF alpha antibody [EPR22598-212] 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 Anti-TNF alpha antibody [EPR22598-212] 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] (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
![Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (ab255288), expandable thumbnail](https://content.abcam.com/products/images/tnf-alpha-antibody-epr22598-212-bsa-and-azide-free-ab255288--flow-cytometry-intracellular-img415544.jpg "Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (ab255288)")
Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [EPR22598-212] - BSA and Azide free (ab255288)
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 Anti-TNF alpha antibody [EPR22598-212] 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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (Anti-TNF alpha antibody [EPR22598-212] ab255275).
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