TNF alpha Rabbit Polyclonal Antibody

• Catalog Number: orb313248
• Category: Antibodies
• Description: TNF alpha Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, WB
• Predicted Reactivity: Canine, Equine, Porcine
• Reactivity: Human, Mouse
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human TNF alpha (86-150/233aa)
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, Flow-Cyt=1ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 17.5/26 kD kDa
• Target: TNF
• UniProt ID: P01375
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Tumor necrosis factor-α; TNFα; TNF-α; TNFalpha; AP
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: Raw264.7. Primary Antibody (green line): Rabbit Anti-TNF alpha antibody (orb313248), dilution: 2 ug/Test, Secondary Antibody: Goat anti-rabbit IgG-FITC, dilution: 0.5 ug/Test. Protocol, The cells were treated with LPS (1 ug/ml, 18 hr/6 hr) and Brefeldin A (300 ng/ml, last 3 hr of stimulation). The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Blank control: Raw264.7. Primary Antibody (green line): Rabbit Anti-TNF alpha antibody (orb313248), dilution: 2 ug/Test, Secondary Antibody: Goat anti-rabbit IgG-FITC, dilution: 0.5 ug/Test. Protocol, The cells were treated with LPS (1 ug/ml, 18 hr/6 hr) and Brefeldin A (300 ng/ml, last 3 hr of stimulation). The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Blank control: THP-1. Primary Antibody (green line): Rabbit Anti-TNF alpha antibody (orb313248), dilution: 1 ug/Test, Secondary Antibody: Goat anti-rabbit IgG-FITC, dilution: 0.5 ug/Test. Protocol, The cells were treated with TPA (80 nM, overnight) and then treated with LPS (1 ug/ml, 18 hr/6 hr) and Brefeldin A (300 ng/ml, last 3 hr of stimulation). The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Sample: Lane 1: A431 (Human) Cell Lysate at 30 ug, Lane 2: A431 (Human) Cell Lysate at 30 ug, Primary: Anti-TNF alpha (orb313248) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 26/17 kD, Observed band size: 26 kD.