TNF Antibody

SKU: orb750677

Description

TNF antibody

Images & Validation

• Tested Applications: ELISA, IF, IHC, WB
• Dilution Range: ELISA: 1:1,000 - 1:5,000, IHC: 1:100 - 1:500, IF: 1:100, WB: 1:500 - 1:2,000
• Reactivity: Human, Primate
• 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.

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Antiserum
• Immunogen: The whole rabbit serum was prepared by repeated immunizations with recombinant human TNFa produced in E.coli.
• Target: TNF
• 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

Storage & Handling

• Storage: Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.
• Form/Appearance: Liquid (sterile filtered)
• Buffer/Preservatives: None. None
• Concentration: 75 mg/mL
• Expiration Date: 12 months from date of receipt.
• Dry Ice Shipping: Please note: This product requires shipment on dry ice. A dry ice surcharge will apply.
• Disclaimer: For research use only

Alternative Names

APC1 antibody, Cachectin antibody, DIF antibody, Differentiation inducing factor antibody, Macrophage cytotoxic factor antibody, MCF antibody, Necrosin antibody, Tumour Necrosis Factor Alpha antibody, rabbit anti-Tumor Necrosis Factor Alpha Antibody, rabbit anti-TNF Alpha Antibody, Tumor necrosis factor, TNF-alpha, Tumor necrosis factor ligand superfamily member 2, TNF-a, Tumor necrosis factor membrane form, N-terminal fragment, NTF, Intracellular domain 1, ICD1, Intracellular domain 2, ICD2, C-domain 1, C-domain 2, Tumor necrosis factor, soluble form, TNF, TNFA, TNFSF2

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 (pH6.0). 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: 500ng 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.

Quick Database Links

Gene Symbol

TNF

UniProt

P01375

RefSeq:P01375.1