TNF alpha antibody

• Catalog Number: orb420293
• Category: Antibodies
• Description: TNF alpha antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IF, IHC, WB
• Reactivity: Human
• Isotype: IgG
• Immunogen: TNF alpha antibody used to produce this IgG fraction antibody was prepared by repeated immunizations with recombinant human TNFα.
• Concentration: 1.0 mg/ml
• Dilution range: ELISA: 1:1,000 - 1:5,000, IHC: 1:100 - 1:500, IF: User Optimized, WB: 1:500 - 1:2,000
• Form/Appearance: Liquid (sterile filtered)
• 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
• UniProt ID: P01375
• NCBI: P01375.1
• Storage: Store anti-TNF alpha 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
• Alternative names: APC1 antibody, Cachectin antibody, DIF antibody, D
• Note: For research use only
• 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.
• 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 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: 26 kDa 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.