TNF Antibody

SKU: orb345126

Description

TNF Alpha antibody

Images & Validation

• Tested Applications: ELISA, IF, IHC, WB
• Dilution Range: ELISA: 1:1,000 - 1:5,000, IHC: 1:100 - 1:500, WB: 1:500 - 1:2,000
• Reactivity: Human
• Application Notes:
  This IgG fraction antibody of anti-Human TNFα has been tested for use in neutralizations, ELISA, immunohistochemistry 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: IgG
• Immunogen: The whole rabbit serum used to produce this IgG fraction antibody was prepared by repeated immunizations with recombinant human TNFα.
• Target: TNF
• Purity: This antibody is primarily directed against mature 17,000 MW human TNFα and is useful in determining its presence in various assays. In general, this antibody also detects primate TNFα in the same formats using similar dilutions. The antibody does not recognize human TNFß (lymphotoxin). This IgG fraction antibody will recognize the cell-bound precursor of TNFα as a 26,000 protein in immunoblots, particularly in denatured samples. This antibody is also useful for neutralization of human and primate TNFα 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: Preservative: None. Stabilizer: None; Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Concentration: 1.0 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

(I) Immunostaining for TNF-a: (A) negative control; (B) muscle from subject 465 yr (positive control); (C) muscle from weight maintainer; and (D) muscle from weight gainer. Anti-human TNF-a antibody (p/n orb345125) at 1:50. The sites of peroxidase binding were demonstrated with diamonobenzidine.

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 Rabbit anti-TNF Alpha antibody. Marker: Opal Pre-stained ladder. Lane 1: HEK293 lysate (p/n orb348669). Lane 2: HeLa Lysate (p/n orb348668). Lane 3: MCF-7 Lysate (p/n orb348664). Lane 4: Jurkat Lysate. Lane 5: A431 Lysate (p/n orb348665). Lane 6: A549 Lysate (p/n orb348675). Lane 7: LNCap Lysate (p/n orb348694). Lane 8: MOLT-4 Lysate (p/n orb348696). Lane 9: Ramos Lysate. Lane 10: Raji Lysate (p/n orb348672). Lane 11: A-172 Lysate (p/n orb348708). Lane 12: NIH/3T3 Lysate (p/n orb348714). Load: 35 µg per lane. Primary antibody: TNF Alpha antibody at 1 ug/ml overnight at 4C. Secondary antibody: Peroxidase rabbit secondary antibody (p/n orb347654) at 1:30000 for 60 min at RT. Blocking Buffer: 1% Casein-TTBS for 30 min at RT. Predicted/Observed size: 26kDa for TNF Alpha.

Western blot using Biorbyt's Anti-Human TNF-a (RABBIT) Antibody. Membrane blocked in 1% BSA-TBS-T for 30 min at 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 buffer (p/n orb348637) for 30 min at RT.

Quick Database Links

Gene Symbol

TNF

UniProt

P01375

RefSeq:P01375.1