Anti-TLR4抗体[HTA125] (ab8376)

Thank you for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results. I apologize for the inconvenience and am pleased to offer you a credit note/ refund in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

PS: Please note as the refund or credit note will be given so theimage discount code provided for this antibody will not be valid.

Thank you for your email.

I have checked the images and based on the images and protocol, I have following suggestions;

- Use a different HRP conjugated antibody. DAKO ChemMate EnVision Detection Kit as per DAKO website is optimized for IHC only. You might need to check whether the antibody DAKO ab is suitable for western blot. Please check with DAKO also.

- Try heating the lysates 70C for 15-20 minutes. As TLR4 is a single pass membrane protein so heating at low incubation temperature for longer period of time would help.

- The background staining can be reduced using 5% BSA as an alternative to milk. Please try BSA.

- Please check and optimize the tissue homogenization/ protein preparation protocol.

- Please use RAW or Daudi cell lysates as positive control or cell lysates other than the TCA prepared lysates.

I hope these suggestions would help. Should the results do not improve please let me know I will be happy to replace the antibody.

Thank you very much sending details.

I am surprised to hear that antibody failed in 2 different occasions as we haven't received any complaint for these lots so I think protocol might need further optimizations. Could you provide image of staining? Could you provide how you have prepared the samples and how old the samples were?

I will look forward to hearing from you soon.

Thank you for completing the survey on Abcam website.

I am sorry to hear that the antibody did not work with porcine sample. I have attached a questionnaire, to follow up for the complaint. Could you complete this and send it to me asap. I will then check if troubleshooting of protocol could help.

Many thanks for your cooperation in advance. I will look forward to hearing from you soon.

Thank you for your email.

The order number I have created is 1107878; you will be receiving this antibody shortly.

TLR4 is highly expressed in porcine kidney so the antibody should have worked. Please let me know if you experience any problem again.

Could I recommend using human, kidney, liver, placenta or spleen tissue lysates as positive control lysates, just to double check if the porcine lysates are OK.

Thank you for contacting us.

I am sorry, I forgot to ask the order number of your last purchase. Could you send the purchase order number to us so that we can send the new vial at same address?

Thanks! Looking forward to hearing from you soon.

DISCOUNT CODE: XXXXXXXXXXX (for ab8376)
DISCOUNT CODE: XXXXXXXXXXX (for ab11354)

Expiration date: 24-09-2012

I am very pleased to hear you would like to use ab8376 (WB) and ab11354(IHC-P) inPorcine and send us an image of your results.

This code will give you2 free [PRIMARY ANTIBODY/PROTEIN] before the expiration date. To redeem this offer, please submit an Abreview with the image for [IHC-P and WB] and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

The code will be active once the Abreview with the image has been submitted and can be redeemed in one of the following ways:
1) Call to place your order and mention the code to our customer service department;
2) Include the code in your fax order;
3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated. Remember if the product does not work you will be covered by our Abpromise guarantee and eligible for a full refund or replacement.

If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Thank you for your reply. I'm afraid all the information we have is from those references and therefore we do not have any more data looking at the blocking ability of the antibody. We welcome all feedback on our antibodies via our Abreviews system and I hope you will consider sharing your data with other users of the antibody; all feedback is rewarded with Abpoints which can be used for gifts and discounts. I'm sorry I couldn't help you more on this occasion and wish you all the best with your research,

I can confirm that the blocking activity of this clone has been established in a number of studies, for your convenience I have listed some useful articles below: 1: Infect Immun. 2001 Apr;69(4):2045-53. Synergistic effect of muramyldipeptide with lipopolysaccharide or lipoteichoic acid to induce inflammatory cytokines in human monocytic cells in culture. Yang S, Tamai R, Akashi S, Takeuchi O, Akira S, Sugawara S, Takada H. 2: J Exp Med. 2002 Jan 7;195(1):99-111. Oligosaccharides of Hyaluronan activate dendritic cells via toll-like receptor 4. Termeer C, Benedix F, Sleeman J, Fieber C, Voith U, Ahrens T, Miyake K, Freudenberg M, Galanos C, Simon JC. 3: Infect Immun. 2001 Apr;69(4):2402-6. Involvement of CD14 and toll-like receptors in activation of human monocytes by Aspergillus fumigatus hyphae. Wang JE, Warris A, Ellingsen EA, Jorgensen PF, Flo TH, Espevik T, Solberg R, Verweij PE, Aasen AO. 4: J Immunol. 2000 Jul 1;165(1):411-8. Proteolysis of human monocyte CD14 by cysteine proteinases (gingipains) from Porphyromonas gingivalis leading to lipopolysaccharide hyporesponsiveness. Sugawara S, Nemoto E, Tada H, Miyake K, Imamura T, Takada H. 5: J Exp Med. 1999 Jun 7;189(11):1777-82. MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-likereceptor 4. Shimazu R, Akashi S, Ogata H, Nagai Y, Fukudome K, Miyake K, Kimoto M. Please note that the references and datasheet mention the clone HTA125, which should be used Azide free (or using ab30667 which is supplied with no Azide). I wonder therefore if the lack of blocking you observe could therefore be due to the presence of Azide. I hope the above references will help you,

Unfortunately I do not believe that the epitope that this antibody recognises has been fully characterised and I have been unable to locate any further information relating to the terminus recognised. I am sorry that I cannot be of additional assistance in this matter,