Recombinant human TNF alpha protein (Active) (ab259410)

Recombinant human TNF alpha protein (Active) is a Human Full Length protein in the 77 to 233 aa range with >=95% purity, < 0.005 EU/µg endotoxin level and suitable for Functional studies, Cell Culture, ELISA, SDS-PAGE and more. The predicted molecular weight of ab259410 protein is 17.4 kDa.

- Save time and ensure accurate results- use our TNF alpha protein as a control
- Optimal protein bioactivity and stability
- Available in different sizes to fit your experimental needs

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Images

Flow Cytometry - Recombinant human TNF alpha protein (Active) (AB259410), expandable thumbnail

Publications

Key facts

Purity: >=95% SDS-PAGE
Endotoxin level: < 0.005 EU/µg
Expression system: HEK 293 cells
Tags: Tag free
Applications: SDS-PAGE, FuncS, MS, sELISA, Cell Culture, HPLC
Biologically active: Yes

Amino acid sequence

V R S S S R T P S D K P V A H V V A N P Q A E G Q L Q W L N R R A N A L L A N G V E L R D N Q L V V P S E G L Y L I Y S Q V L F K G Q G C P S T H V L L T H T I S R I A V S Y Q T K V N L L S A I K S P C Q R E T P E G A E A K P W Y E P I Y L G G V F Q L E K G D R L S A E I N R P D Y L D F A E S G Q V Y F G I I A L

Reactivity data

Application	Reactivity	Dilution info	Notes
Application SDS-PAGE	Reactivity Reacts	Dilution info -	Notes -
Application FuncS	Reactivity Reacts	Dilution info -	Notes -
Application MS	Reactivity Reacts	Dilution info -	Notes -
Application sELISA	Reactivity Reacts	Dilution info -	Notes -
Application Cell Culture	Reactivity Reacts	Dilution info -	Notes -
Application HPLC	Reactivity Reacts	Dilution info -	Notes -

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Target data

See full target information TNF

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

Key facts

Purity: >=95% SDS-PAGE
Endotoxin level: < 0.005 EU/µg
Expression system: HEK 293 cells
Tags: Tag free
Applications: SDS-PAGE, FuncS, MS, sELISA, Cell Culture, HPLC
Biologically active: Yes
Biological activity: Fully biologically active when compared to standard. The ED₅₀ as determined by the dose-dependant Killing/apoptosis of L-929 cells is 0.71ng/mL corresponding to a Specific Activity of 1.41 x 10⁶ IU/mg.
Accession: P01375-1
Animal free: Yes
Species: Human
Reconstitution: Reconstitute in PBS
Concentration
Storage buffer: pH: 6 - 8
Constituents: 10.26% Trehalose, 0.727% Dibasic monohydrogen potassium phosphate, 0.248% Potassium phosphate monobasic

Sequence info

Amino acid sequence

V R S S S R T P S D K P V A H V V A N P Q A E G Q L Q W L N R R A N A L L A N G V E L R D N Q L V V P S E G L Y L I Y S Q V L F K G Q G C P S T H V L L T H T I S R I A V S Y Q T K V N L L S A I K S P C Q R E T P E G A E A K P W Y E P I Y L G G V F Q L E K G D R L S A E I N R P D Y L D F A E S G Q V Y F G I I A L
Accession: P01375
Protein length: Full Length
Predicted molecular weight: 17.41 kDa
Amino acids: 77 to 233
Nature: Recombinant

Specifications

Form: Lyophilized
Additional notes: >= 95 % HPLC.

General info

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.
Sequence similarities: Belongs to the tumor necrosis factor family.
Post-translational modifications: The soluble form derives from the membrane form by proteolytic processing. The membrane-bound form is further proteolytically processed by SPPL2A or SPPL2B through regulated intramembrane proteolysis producing TNF intracellular domains (ICD1 and ICD2) released in the cytosol and TNF C-domain 1 and C-domain 2 secreted into the extracellular space.

Storage

Shipped at conditions: Ambient - Can Ship with Ice
Appropriate short-term storage conditions: Ambient
Appropriate long-term storage conditions: Ambient

This product is an active protein and may elicit a biological response in vivo, handle with caution.

Notes

Ensure the validity of your result using our recombinant human TNF alpha protein ab259410 as a control.

The ab259410 TNF alpha protein is sourced from HEK293 cells and can be used as a positive control in SDS-PAGE, mass spectrometry and HPLC. Analyze your TNF alpha ELISA data using the ab259410 protein to generate and plot a standard curve.

Check out our protein gel staining guide for SDS-PAGE here

Check out our ELISA protocol for more information here.

Premium Bioactive Protein range

The ab259410 TNF alpha protein is part of the premium bioactive protein range which are ideal for preclinical cell culture and functional studies. These recombinant proteins are of the highest activity, purity, and consistency, meeting rigorous biophysical characterization.

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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.

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7 product images

Flow Cytometry - Recombinant human TNF alpha protein (Active) (ab259410)
Flow cytometry overlay histogram showing wild-type A549 (green line) and VCAM1 knockout A549 cells (red line, Human VCAM1 knockout A549 cell line ab273758), treated with 10 ng/ml TNF-alpha for 16 h (left) and untreated (right), stained with APC Anti-VCAM1 antibody [STA] ab103173. The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (APC Anti-VCAM1 antibody [STA] ab103173) (1x10⁶ in 100μl at 0.2μg/ml) for 30 min at 4°C.
Isotype control antibody mouse IgG1κ Allophycocyanin was used at the same concentration and conditions as the primary antibody (wild-type A549 - black line VCAM knockout A549 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.
Sandwich ELISA - Recombinant human TNF alpha protein (Active) (ab259410)
Sandwich ELISA - Recombinant human TNF alpha protein standard curve.
Background subtracted standard curve using Human TNF alpha Antibody Pair - BSA and Azide free (Human/Monkey TNF alpha Antibody Pair - BSA and Azide free ab241791) and Recombinant human TNF alpha protein (Active) (ab259410) in sandwich ELISA. The ELISA was performed using the components of the corresponding SimpleStep® kit, which uses the same antibody pair with a different formulation and format.
Functional Studies - Recombinant human TNF alpha protein (Active) (ab259410)
Fully biologically active when compared to standard. The ED₅₀ as determined by the dose-dependant Killing/apoptosis of L-929 cells is 0.71 ng/mL corresponding to a Specific Activity of 1.41 x 10⁶ IU/mg.
Sandwich ELISA - Recombinant human TNF alpha protein (Active) (ab259410)
Wild-type A549 control cells or IP-10 knockout A549 cells (Human CXCL10 (IP10) knockout A549 cell line ab266969), grown to 40% confluency, were stimulated with Recombinant Human Interferon gamma protein (Recombinant Human Interferon gamma protein (Active) ab259377) at 100 ng/ml and Recombinant human TNF alpha protein (ab259410) at 10 ng/ml or vehicle control for 16 or 32 hours.
THP-1 cells, grown to 40% confluency, were stimulated with Recombinant Human Interferon gamma protein (Recombinant Human Interferon gamma protein (Active) ab259377) at 200 ng/ml and LPS at 50 ng/mL or vehicle control for 24 hours.
The concentrations of IP-10 (CXCL10) in cell culture supernatants were measured in duplicate and interpolated from the IP-10 standard curves using Human IP-10 ELISA Kit (Human IP-10 ELISA Kit ab173194) . IP-10 from vehicle control samples were measured in undiluted supernatants and the treated samples were diluted 200 times. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
SDS-PAGE - Recombinant human TNF alpha protein (Active) (ab259410)
SDS-PAGE analysis of ab259410.
HPLC - Recombinant human TNF alpha protein (Active) (ab259410)
Purity: 100%
The spectrum was recorded using a 1260 Infinity II HPLC system with DAD and a MabPac RP column (3.0x100 mm, 4 μm). 5 μL of purified protein was injected and the gradient run from 80 % water:TFA (99.9:0.1 v/v) and 20 % acetonitrile:water:TFA (90:9.9:0.1 v/v/v) to 20 % water:TFA (99.9:0.1 v/v) and 80 % acetonitrile:water:TFA (90:9.9:0.1 v/v/v) within 3 minutes followed by an isocratic step for another 3 min. Flow rate was 0.5 mL/min and the column compartment temperature was 50 °C.
Mass Spectrometry - Recombinant human TNF alpha protein (Active) (ab259410)
M + 0.2 Da (calc. mass 17409.8 Da)
The spectrum was recorded with a 6545XT AdvanceBio LC/Q-TOF (Agilent Technologies) and a MabPac RP column (42.1x50 mm, 4 μm, Thermo Scientific). 5 μL of purified protein was injected and the gradient run from 85 % water:FA (99.9:0.1 v/v) and 15 % acetonitrile:FA (90:9.9:0.1 v/v/v) to 55 % water:FA (99.9:0.1 v/v) and 45 % acetonitrile:FA (90:9.9:0.1 v/v/v) within 3 minutes followed by an isocratic step for another 2.5 min. Flow rate was 0.4 mL/min and the column compartment temperature was 60 °C. Data was analysed and deconvoluted using the Bioconfirm software (Agilent Technologies).