TNF antibody

• Catalog Number: orb750676
• Category: Antibodies
• Description: TNF antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IF, IHC, WB
• Reactivity: Human, Primate
• Isotype: Antiserum
• Immunogen: The whole rabbit serum was prepared by repeated immunizations with recombinant human TNFa produced in E.coli.
• Concentration: 75 mg/ml
• Dilution range: ELISA: 1:1,000 - 1:5,000, IHC: 1:100 - 1:500, IF: 1:100, WB: 1:500 - 1:2,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: This antiserum has been heated to 56°C for 30 minutes. The antiserum is directed against mature 17,000 MW human TNFa and is useful in determining its presence in various assays. In general, this antibody also detects primate TNFa in the same formats using similar dilutions. The antibody does not recognize human TNFb (lymphotoxin). This antiserum will recognize the cell-bound precursor of TNFa as a 26,000 protein in immunoblots, particularly in denatured samples. This antibody is also useful for neutralization of human and primate TNFa activity in bioassays. It does not neutralize the biological activity of lymphotoxin. For neutralization, it is recommended to incubate the sample with a 1:200 dilution of the antibody for at least 4 hours before being tested. A control of similarly diluted normal rabbit IgG is recommended.
• Conjugation: Unconjugated
• UniProt ID: P01375
• NCBI: P01375.1
• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: None. None
• Alternative names: APC1 antibody, Cachectin antibody, DIF antibody, D
• Note: For research use only
• Application notes: Anti-Human TNFα has been tested for use in immunohistochemistry, immunofluorescence, and immunoblotting. It recognizes the 17,000 MW TNFα. Reactivity in other immunoassays is unknown.
• Expiration Date: 12 months from date of receipt.

Fluorescent immunohistochemistry showing staining of human colon by Biorbyt's anti-TNF alpha (formalin/PFA-fixed paraffin-embedded sections). Samples were formaldehyde-fixed, then blocked in 10% serum for 2 hours at 20°C. The primary antibody was diluted 1:100 and incubated with the sample for 2 hours at 20°C. Alexa Fluor® 680 goat polyclonal secondary antibody was used diluted 1:5000.

Immunohistochemistry using Biorbyt's polyclonal TNFa antibody showing staining of formalin/PFA-fixed paraffin-embedded sections of human artery tissue sections. Sections were fixed in formaldehyde and subjected to heat mediated antigen retrieval in citrate buffer (pH 6.0). Slides were blocked for ten minutes with 1.5% serum. Primary antibody was diluted 1:100 and incubated with samples for 24 hours at 4°C. HRP-conjugated goat anti-rabbit antibody was used as the secondary antibody.

Western blot of Anti-Human TNF-a (RABBIT) Antibody. Lane 1: 250 ng human recombinant TNF. Lane 2: 500 ng human recombinant TNF. Lane 3: 1000 ng human recombinant TNF. Membrane blocked in Blotto (p/n orb348624) for 30 min RT. Primary antibody: Rb-a-TNF alpha added at 1:1000 in overnight at 4°C, Gt-a-Rb HRP (p/n orb347654) added at 1:20000 in 5% Blotto for 30 min RT. (5 second exposure).

Western blot using Biorbyt's Anti-Human TNF-a (RABBIT) Antibody. Membrane blocked in 1% BSA-TBS-T 30 min RT, Rb-a-TNF alpha added at 1:1000 in 1% BSA-TBS-T o/n 4°C, DyLight 649 Gt-a-Rb added at 1:20000 in (p/n orb348637) 30 min RT.