Overview

  • Product name
    Anti-IL-8 antibody [807]
    See all IL-8 primary antibodies
  • Description
    Mouse monoclonal [807] to IL-8
  • Host species
    Mouse
  • Specificity
    This antibody recognizes both, recombinant and native human IL-8. This antibody does not cross react with human Monocyte Chemotactic Activating Factor (MCAF) or RANTES (Regulated on Activation, Normal T-cell Expressed, and Secreted).
  • Tested applications
    Suitable for: ELISA, Neutralising, WB, IHC-Fr, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human, Pig
  • Immunogen

    Purified recombinant human IL-8.

  • Positive control

Properties

Applications

Our Abpromise guarantee covers the use of ab18672 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ELISA Use at an assay dependent concentration.
Neutralising Use at an assay dependent concentration.

In chemotaxis assay, the antibody inhibited 95.1% of the chemotactic activity of IL-8 on RB/293 cell. 50ug/ml of antibody and 10ng/ml of human IL-8 were used in assay. This antibody inhibited the chemotactic effect of human IL-8, and had no inhibitory effect of MIP 1-beta and RANTES in neutrophil chemotaxis assay, indicating that the neutralizing activity is specific. The antibody also neutralized IL-8 induced calcium influx in human granulocytes.

WB Use a concentration of 0.02 - 0.1 µg/ml. Predicted molecular weight: 12 kDa.

Concentration of 0.02-0.1 ug/ml of ab18672 will allow visualization of 100 ng/lane of human IL-8

IHC-Fr 1/2000.
IHC-P 1/2000.
ICC/IF Use at an assay dependent concentration. PubMed: 18697197

Target

  • Function
    IL-8 is a chemotactic factor that attracts neutrophils, basophils, and T-cells, but not monocytes. It is also involved in neutrophil activation. It is released from several cell types in response to an inflammatory stimulus. IL-8(6-77) has a 5-10-fold higher activity on neutrophil activation, IL-8(5-77) has increased activity on neutrophil activation and IL-8(7-77) has a higher affinity to receptors CXCR1 and CXCR2 as compared to IL-8(1-77), respectively.
  • Sequence similarities
    Belongs to the intercrine alpha (chemokine CxC) family.
  • Post-translational
    modifications
    Several N-terminal processed forms are produced by proteolytic cleavage after secretion from at least peripheral blood monocytes, leukcocytes and endothelial cells. In general, IL-8(1-77) is referred to as interleukin-8. IL-8(6-77) is the most promiment form.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • (Ala-IL-8)77 antibody
    • (Ser-IL-8)72 antibody
    • 9E3 antibody
    • Beta thromboglobulin like protein antibody
    • C-X-C motif chemokine 8 antibody
    • CEF-4 antibody
    • chemokine, CXC motif, ligand 8 antibody
    • CXCL8 antibody
    • Emoctakin antibody
    • GCP-1 antibody
    • GCP/IL-8 protein I antibody
    • GCP/IL-8 protein II antibody
    • GCP/IL-8 protein III antibody
    • GCP/IL-8 protein IV antibody
    • GCP/IL-8 protein V antibody
    • GCP/IL-8 protein VI antibody
    • GCP1 antibody
    • Granulocyte chemotactic protein 1 antibody
    • IL-8 antibody
    • IL-8(1-77) antibody
    • IL-8(9-77) antibody
    • IL8 antibody
    • IL8/NAP1 form I antibody
    • IL8/NAP1 form II antibody
    • IL8/NAP1 form III antibody
    • IL8/NAP1 form IV antibody
    • IL8/NAP1 form V antibody
    • IL8/NAP1 form VI antibody
    • IL8_HUMAN antibody
    • Inteleukin 8 antibody
    • LECT antibody
    • LUCT antibody
    • Lymphocyte-derived neutrophil-activating factor antibody
    • LYNAP antibody
    • MDNCF antibody
    • MDNCF-b antibody
    • MDNCF-c antibody
    • MONAP antibody
    • Monocyte-derived neutrophil chemotactic factor antibody
    • Monocyte-derived neutrophil-activating peptide antibody
    • NAF antibody
    • NAP-1 antibody
    • NAP1 antibody
    • Neutrophil activating peptide 1 antibody
    • Neutrophil activating protein 1 antibody
    • Neutrophil-activating factor antibody
    • Neutrophil-activating protein 1 antibody
    • Protein 3 10C antibody
    • Protein 3-10C antibody
    • SCYB8 antibody
    • Small inducible cytokine subfamily B member 8 antibody
    • T cell chemotactic factor antibody
    • T-cell chemotactic factor antibody
    • TSG1 antibody
    see all

Images

  • ab18672 staining IL8 in porcine epithelial tissue by Immunohistochemistry (Frozen sections). Tissue was fixed in paraformaldehyde, permeabilized using 0.25% Triton X-100, blocked with 10% serum for 2 hours at 25°C, then incubated with ab18672 at a 1/2000 dilution for 12 hours at 4°C. The secondary used was ab98640, used at a 1/1000 dilution.

    See Abreview

  • ab18672 staining IL8 in inflammed human temporal artery by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    Tissue was fixed in formaldehyde and an antigen retrieval step was performed using sodium citrate. Samples were then blocked with 5% serum for 1 hour at 25°C, then incubated with ab18672 at a 1/2000 dilution for 16 hours at 4°C. The secondary used was a rabbit anti-mouse polyclonal conjugated to biotin, 1/1000 dilution.

    See Abreview

  • Anti-IL-8 antibody [807] (ab18672) at 0.1 µg/ml + recombinant human IL8, 100 ng

    Predicted band size: 12 kDa

References

This product has been referenced in:
  • Tengroth L  et al. Activation of Activin receptor-like kinases curbs mucosal inflammation and proliferation in chronic rhinosinusitis with nasal polyps. Sci Rep 8:1561 (2018). Read more (PubMed: 29367682) »
  • Wadghiri YZ  et al. High-resolution Imaging of Myeloperoxidase Activity Sensors in Human Cerebrovascular Disease. Sci Rep 8:7687 (2018). Read more (PubMed: 29769642) »
See all 12 Publications for this product

Customer reviews and Q&As

1-10 of 21 Abreviews or Q&A

Application
ELISA
Sample
Human Recombinant protein (Human IL-8 from R&D Systems)
Specification
Human IL-8 from R&D Systems
Type
Sandwich (Capture)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Mar 07 2014

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Sample
Human Tissue sections (tumor)
Specification
tumor
Permeabilization
Yes - PBS+0.3% Triton-X-100
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Dec 06 2013

Answer

Thank you for contacting us.
1. If you prefer to do the incubation at room temperature you should not exceed a 2 hour incubation. We do recommend an overnight incubation at 4 degrees, however, as this usually gives the strongest and most specific staining.
2. I would suggest you use ab6672 at 1/400 and ab18672 at 1/2000.
3. I believe lung cancer tissue is a good positive control for these antibodies but am not sure that the levels in normal tissue are high enough. For ab18672 we have seen good expression in lymph node, poocine epithelial tissue, and human temporal artery. For ab6672 human and mouse pancreatic cancer, head and neck tumor, and human liver sections have shown good results. Any tissue reported in the literature to have good IL6 or IL8 expression should be a good positive control if you do not have access to these tissues.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your reply.

I am sorry that this antibody did not perform as stated on the datasheet. As requested, I have asked our Finance department to issue a credit note for you

Credit note ID: 23605

The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

The credit note is valid for 15 months.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

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Answer

Thank you for your reply.

I fully understand your concerns regarding the product code typing error on the datasheet, it is regrettable. I would like to reassure you that the image is from ab18672 IL8 antibody [807], and came from an Abreview submitted by a customer which is published on the Abreview page of the datasheet:

https://www.abcam.com/index.html?datasheet=18672&tab=abreviews&intabreviewid=24612

I can confirm that any publications that we are aware of will be listed on the datasheet. We have a dedicated team who spend time reviewing the literature to find articles that have used our products. However, in many cases, regrettably we do not have access to all the articles mentioning our products, or subscriptions to all the journals that publish them. I'm sorry there are no others to provide on this occasion for ab18672.

Regarding the product quality, I would like to reassure you that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this batch. Regrettably, I can suggest you have received a bad vial on this occasion. Therefore, we would be pleased to provide a replacement from the same antibody, which will still be covered by our guarantee. Alternatively, I fully understand your concerns and if you prefer an alternative replacement or credit note in this case, I will be pleased to arrange this for you.

I hope this will be helpful. I look forward to hearing from you with details of how you would like to proceed.

Read More

Answer

Thank you for your message and for providing this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments, and would like to offer a free of charge replacement or credit note in compensation, providing the product has been used in a tested and guaranteed species. Before arranging this, I would appreciate if you could confirm which species the samples are from?

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

Read More

Question
Answer

Thank you for your enquiry.

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

With regards to the text with the image, Ihave investigated this with my colleagues from our datasheetsteam and I can confirm that regrettably there was a typing error. I can confirm this IHC image is fromab18672 Anti-IL8 antibody [807].The datasheet has been updated with the correct information and I apologize for any confusion.

The quality of our antibodies and datasheets is very important to us, as is any feedback and data we receive. It is regrettable that the information we had with the image on this datasheet was not of the standard both we and our customers expect from our products.

With regards to the difficulty with staining, I would like to reassure you that this antibody is tested and covered by our 6 month guarantee forIHC-P and in Human andPig samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if you are also able to provide an image which would help us to assess the results.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.



Order Details
Antibody code:

Lot number:


Purchase order number
or preferably Abcam order number:


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, low signal, non-specific satining etc.)

Sample (Species/Tissue/Cell Type/Cell Line etc.)


Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)


Antigen retrieval (Enzymatic method, Heat mediated technique etc.)


Permeabilization step


Blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method


Positive and negative controls used (please specify)


Optimization attempts (problem solving)
How many times have you tried the IHC?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No


What steps have you altered?


Additional Notes


We would appreciate if you are also able to provide and image which woudl help us to assess the results

Read More

Answer

Thank you for contacting us.

Unfortunately there is no information available about the epitope recognised by these two antibodies, as it hasn’t been mapped.

I am very sorry I couldn’t help you further. Please do not hesitate to contact us if you need any other advice or information.

Read More

Answer

Thank you again for your call last week and for your patience while I have reviewed your images and these proteins.

For ab18672 (IL8), I think the diffuse bands are caused bya bittoo much proteinloaded in the lanes orfrom running the gel at too high a voltage. You can try to load a bit less protein and lower the voltage during electrophoresis. It's also possible that the two isoforms of IL8 are being expressed and since they're so close in MW, they are resulting in a thicker band, but I would still try the other alterations to see if they tighten the bands.

Ab24598 (thrombomodulin) is probably picking up glycosylated forms of the protein, which has 7 known glycosylation sites. Each glycosylation can add 2 kDa to the predicted molecular weight of the amino acid sequence (around 60 kDa). There may be differentially glycosylated forms of the TM in your sample. I found a reference showing a similar doublet around this MW as well (see attached).

The results with thePTG antibody, ab103300, area little harder to analysis. I've looked through a lot of literature, but there are not many Western blots showing the endogenous molecular weight. Generally the protein shows up between 35 and 38 kDa, and the variance could be caused by different electrophoretic running conditions, etc, and not actual differences in the protein. Based on the information I've found, the higher band seems to be specific and the lower band is probably non-specific. According to my notes, you had tried using 5 ug/mL with this antibody, so I would suggest decreasing the amount of antibody used to reduce any non-specific binding. This should help determine the specificity of the higher band.

I hope that this information will be helpful, and if you have any questions or further comments please let me know. Please keep me updated about any new results with these antibodies. If the results are not satisfactory at any time, these antibodies are covered by our Abpromise guarantee and we can arrange a replacement, credit, or refund.

Have a nice day.

Read More

Answer

Thank you very much for your call today and for letting us know about the trouble with these antibodies.

As we discussed, please send any images of the blots and we will see if there are some suggestions to improve the results. Could you also tell me what percentage of gel was used?

I look forward to hearing from you. Please let me know if you have further questions or if there is anything else that we can do for you.

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1-10 of 21 Abreviews or Q&A

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