TNF-alpha Monoclonal antibody

• Featured Product
• KD/KO Validated

TNF-alpha Monoclonal Antibody for WB, IHC, IF-P, ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

human, rat and More (7)

Applications

WB, IHC, IF-P, RIP, ELISA

Conjugate

Unconjugated

Publications

489

CloneNo.

7B8A11

Reviews (2)

Cat no : 60291-1-Ig

Synonyms

TNF, TNF alpha, TNF-a, Intracellular domain 2, Intracellular domain 1

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Validation Data Gallery

• 

  WB analysis using 60291-1-Ig

  PMA, LPS, and Brefeldin A treated THP-1 cells were subjected to SDS PAGE followed by western blot with 60291-1-Ig (TNF Alpha antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with GAPDH Monoclonal antibody 60004-1-Ig as loading control.

• 

  WB analysis of HeLa using 60291-1-Ig

  WB result of TNF alpha antibody (60291-1-Ig; 1:2000; incubated at room temperature for 1.5 hours) with sh-Control and sh-TNF alpha transfected HeLa cells.

• 

  WB analysis using 60291-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 60291-1-Ig (TNF Alpha antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 60291-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 60291-1-Ig (TNF alpha antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HeLa using 60291-1-Ig

  HeLa cells were subjected to SDS PAGE followed by western blot with 60291-1-Ig (TNF alpha antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HL-60 using 60291-1-Ig

  HL-60 cells were subjected to SDS PAGE followed by western blot with 60291-1-Ig (TNF alpha antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of U-937 using 60291-1-Ig

  U-937 cells were subjected to SDS PAGE followed by western blot with 60291-1-Ig (TNF alpha antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 60291-1-Ig

  Untreted and PMA treated THP-1 cells were subjected to SDS PAGE followed by western blot with 60291-1-Ig (TNF Alpha antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HeLa using 60291-1-Ig

  HeLa cells were subjected to SDS PAGE followed by western blot with 60291-1-Ig (TNF Alpha antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of human breast cancer using 60291-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 60291-1-Ig (TNF Alpha antibody) at dilution of 1:400 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 60291-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 60291-1-Ig (TNF Alpha antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human tonsillitis using 60291-1-Ig

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 60291-1-Ig (TNF Alpha antibody) at dilution of 1:800 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human tonsillitis using 60291-1-Ig

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 60291-1-Ig (TNF Alpha antibody) at dilution of 1:800 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of human breast cancer using 60291-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed human breast cancer tissue using 60291-1-Ig (TNF alpha antibody) at dilution of 1:100 and Alexa Fluor 488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

• 

  IF Staining of human breast cancer using 60291-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed human breast cancer tissue using 60291-1-Ig (TNF alpha antibody) at dilution of 1:100 and Alexa Fluor 488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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Tested Applications

• Positive WB detected in: THP-1 cells, HeLa cells, HL-60 cells, U-937 cells, PC-12 cells, HSC-T6 cells
• Positive IHC detected in: human breast cancer tissue, human tonsillitis tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: human breast cancer tissue

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:4000
• Immunohistochemistry (IHC): IHC : 1:200-1:800
• Immunofluorescence (IF)-P: IF-P : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 1 publications below
• WB: See 340 publications below
• IHC: See 110 publications below
• IF: See 73 publications below
• ELISA: See 4 publications below
• RIP: See 1 publications below

Product Information

60291-1-Ig targets TNF-alpha in WB, IHC, IF-P, RIP, ELISA applications and shows reactivity with human, rat samples.

• Tested Reactivity: human, rat
• Cited Reactivity: human, mouse, rat, pig, rabbit, monkey, chicken, bovine, duck
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: TNF-alpha fusion protein Ag11413
• Full Name: tumor necrosis factor (TNF superfamily, member 2)
• Calculated Molecular Weight: 233 aa, 26 kDa
• Observed Molecular Weight: 17-22 kDa
• GenBank Accession Number: BC028148
• Gene Symbol: TNF-alpha
• Gene ID (NCBI): 7124
• RRID: AB_2833255
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

TNF, as also known as TNF-alpha, or cachectin, is a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. It is expressed as a 26 kDa membrane bound protein and is then cleaved by TNF-alpha converting enzyme (TACE) to release the soluble 17 kDa monomer, which forms homotrimers in circulation. It is produced chiefly by activated macrophages, although it can be produced by many other cell types such as CD4+ lymphocytes, NK cells, neutrophils, mast cells, eosinophils, and neurons. It can bind to, and thus functions through its receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. This cytokine is involved in the regulation of a wide spectrum of biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, INS resistance, and cancer.

Publications

Species	Application	Title
rat	IHC	Signal Transduct Target Ther; Metabolic reprogramming of proinflammatory macrophages by target delivered roburic acid effectively ameliorates rheumatoid arthritis symptoms; Authors - Na Jia
human	IHC	Nat Immunol; NKILA lncRNA promotes tumor immune evasion by sensitizing T cells to activation-induced cell death.; Authors - Di Huang
human	WB	Blood; The hepatocyte-specific HNF4α/miR-122 pathway contributes to iron overload-mediated hepatic inflammation.; Authors - Min Li
human,rat	WB	ACS Nano; Hepatic Stellate Cell- and Liver Microbiome-Specific Delivery System for Dihydrotanshinone I to Ameliorate Liver Fibrosis; Authors - Xia Niu
mouse,rat	WB	Nat Commun; Interventional hydrogel microsphere vaccine as an immune amplifier for activated antitumour immunity after ablation therapy; Authors - Xiaoyu Liu
mouse	WB	Brain Behav Immun; Oxytocin alleviates cognitive and memory impairments by decreasing hippocampal microglial activation and synaptic defects via OXTR/ERK/STAT3 pathway in a mouse model of sepsis-associated encephalopathy; Authors - Junliang Jiang

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Udesh (Verified Customer) (04-19-2023)

Worked well for WB in U2OS cells at 1:1000. TNF-alpha observed in upper membrane part

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: U2OS

TNF-alpha Antibody Western Blot validation (1:1000 dilution) in U2OS (Cat no:60291-1-Ig)

FH

Feng-Qian (Verified Customer) (12-07-2018)

Signal was very weak.

• Applications: Western Blot, Immunofluorescence,
• Primary Antibody Dilution: 1/250
• Cell Tissue Type: HaCaT

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