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Human recombinant tumor necrosis factor of alpha type
Our Abpromise guarantee covers the use of ab8348 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells.|
|Neutralising||Use a concentration of 5 µg/ml.|
|ELISA||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
|IHC-Fr||Use a concentration of 10 - 20 µg/ml.|
|ICC/IF||Use a concentration of 10 µg/ml.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 25 kDa.|
|Sandwich ELISA||Use a concentration of 5 µg/ml. Can be paired for Sandwich ELISA with Rabbit polyclonal to TNF alpha (ab9635). Use this antibody as Capture at 5µg/ml with ab9635 as Detection.|
This image is courtesy of an anonymous Abreview
CON+ = Patient with increased TNF alpha levels on ELISA of muscle homogenate.
POS = Diseased patient with suspected increased TNF alpha levels in muscle.
ab8348 staining TNF alpha in Human skeletal muscle (gastrocnemius) tissue sections by Immunohistochemistry (Formalin/PFA-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% serum for 10 minutes at 25°C; antigen retrieval was by heat mediation in a Tris buffer (pH 8). Samples were incubated with primary antibody (1/200) for 16 hours at 4°C. An undiluted HRP-conjugated Rabbit anti-mouse IgG polyclonal was used as the secondary antibody.
ICC/IF image of ab8348 stained A431 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab8348 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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