Anti-TNF alpha 抗体 (ab9635)

In order to detect Human TNF-α by sandwich ELISA (using 100μl/well antibody solution) a concentration of 0.5 - 2.0 μg/ml of this antibody is required. We recommend determining optimal dilutions of this antibody for your ELISA assay.
The following paper may of use to you for the ELISA protocol used with this antibody:

Our references for Anti-Human TNF-α (500-P31A) include one article citing use in ELISA:
Halin, C, Synergistic therapeutic effects of a tumor targeting antibody fragment, fused to interleukin 12 and to tumor necrosis factor alpha. Cancer Research; 63(12):3202-3210. 15 Jun 2003 (PubMed ID: 12810649).

Thank you for contacting us.

We have in our catalogue the following pairs of antibodies that have been tested for the detection of Human TNF-alpha in sELISA :

Anti-TNF alpha antibody [2C8] (https://www.abcam.com/ab8348) as capture + Rabbit polyclonal to TNF alpha (https://www.abcam.com/ab9635) as detection. Please note that these 2 antibodies are unconjugated.
Anti-TNF alpha antibody [2C8] (https://www.abcam.com/ab8348) as capture + Anti-TNF alpha antibody [F6C5 ] (HRP) (https://www.abcam.com/ab24473) as detection.



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for your inquiry.

We haven't specifically tested ab8348 and ab9635 for cross-reactivity with other proteins.

For ab9635, I BLASTed the immunogen sequence against other human proteins and no significant homology was detected (< 34 %) so we wouldn't anticipate cross-reactivity. Plus this is produced from sera of rabbits pre-immunized with highly pure recombinant Human TNF-α and it was purified by affinity chromatography employing immobilized Human TNF-α matrix.

We have some references for both antibodies that you may be interested in:

https://www.abcam.com/TNF-alpha-antibody-ab9635-references.html

https://www.abcam.com/TNF-alpha-antibody-2C8-ab8348-references.html

The protocol for using the antibodies together in sELISA can be found here: https://www.abcam.com/ps/products/8/ab8348/documents/Protocol%20rabbit.pdf

The ELISA kits we sell may come from different sources, and sometimes we are not provided with the individual antibodies for sale. We can always check individual kits to see if we have the antibodies, but in most cases they probably aren't available.

I hope this information helps. Please contact us with any other questions.

Merci pour votre patience. Veuillez trouver en attaché le document que le laboratoire m'avait envoyé pour le ab9635. Il montre que l'anticorps est efficace à inhiber le TNF alpha à moins que 0.1ug/ml. Veuillez ne pas hésiter à nous recontacter si vous avez besoin de plus d'informations. Je vous souhaite bon succès dans votre recherche.

Thank you for your enquiry and your patience. The neutralization experiment was conducted as follows: Procedure: 1) Resuspend 929 cells in regular culture media. Transfer 100ml/well in assay plate (8,100 cells/well). Incubate 37oC overnight. 2) In separate plate, serial dilute a-h-TNF-a in assay media containing 2mg/ml Actinomycin D and 10% horse serum in triplicate. Add 60ml/well assay media containing 2ng/ml h-TNF-a resulting in 1:2 dilution. Incubate 1 hour. 3) Transfer 100ml/well h-TNF-a/anti-h-TNF-a complex to cells in assay plate Final volume/well is 200ml containing 1mg/ml Actinomycin D, 0.5 ng/ml h-TNF-a and anti-h-TNF-a as indicated below. 4) Incubate 20 hours. 5) Add 20ml of Promega Substrate CellTiter 96 Aqueous One Solution Reagent to each well. 6) Incubate 37oC and read OD at 490 nm. Data: Conc. anti-h-TNF-a O.D. reading at 490nm after 4 hours ug/ml row 1 row 2 row 3 Ave. O.D. Net O.D. 25.00 12.50 6.25 3.13 1.56 0.78 0.39 0.20 0.10 0.00 I hope this information helps, please do not hesitate to contact us if you need any more advice or information,