Overview

  • Product name
  • Description
    Rabbit polyclonal to IL-8
  • Host species
    Rabbit
  • Specificity
    Please be aware that this product has low homology with the mouse and rat sequence of IL8 (Rat, 46%; Mouse 45%, UniProt blast) and we therefore cannot guarantee reactivity in these species.
  • Tested applications
    Suitable for: IHC-P, WB, IHC-Fr, IP, ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Human, Cynomolgus monkey, Rhesus monkey
  • Immunogen

    Recombinant full length protein corresponding to IL-8.

  • Positive control

Properties

Applications

Our Abpromise guarantee covers the use of ab7747 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
IHC-P Use at an assay dependent concentration. PubMed: 17208399
WB 1/10. Detects a band of approximately 6-8 kDa (predicted molecular weight: 11.1 kDa). IL-8 appears to exist as a dimer in solution.
IHC-Fr Use at an assay dependent concentration. PubMed: 25120588
IP 1/10.
ELISA Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 26270987

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

  • ab7747 staining IL8 in SVGA astrocytes transfected with a plasmid encoding Vpr by ICC/IF (Immunocytochemistry/immunofluorescence). The cells were stained for nucleus (blue), IL8 (green) and GFAP (red). Cells were fixed with ice-cold methanol/acetone (1:1) for 20 minutes at 20°C, washed 3x with PBS and permeabilized with 0.1% Triton X-100 in PBS and blocked with 1% BSA for 30 minutes at RT. Samples were incubated with ab7747 overnight in a humidified chamber. The cells were washed 5x with 0.1% Triton X-100 in PBS and incubated for 1h in secondary antibodies conjugated with Alexa Fluor 488 (Anti-Rabbit, 1:1000) at room temperature in the dark. All the antibodies were diluted in PBS containing 1% BSA.

References

This product has been referenced in:
  • Johnson KE  et al. Aspirin inhibits platelets from reprogramming breast tumor cells and promoting metastasis. Blood Adv 3:198-211 (2019). Read more (PubMed: 30670536) »
  • Åström M  et al. Cytokine Measurements for Diagnosing and Characterizing Leukemoid Reactions and Immunohistochemical Validation of a Granulocyte Colony-Stimulating Factor and CXCL8-Producing Renal Cell Carcinoma. Biomark Insights 13:1177271918792246 (2018). Read more (PubMed: 30147294) »
See all 34 Publications for this product

Customer reviews and Q&As

11-14 of 14 Abreviews or Q&A

Answer

That is great to hear. Please contact us again with any more questions or comments.

Read More

Answer

The originator of ab7747 has suggested trying recombinant IL-8 to test your system or using activated macrophages. Since you don't have the original vial, do you have the Abcam order number or the purchase order number? Do you happen to know the lot number of the previous batch that you received which worked for you?

Read More

Question

ANTIBODY CODE ab7747 BATCH NUMBER not sure ORDER NUMBER not sure DESCRIPTION OF THE PROBLEM No signal detected by 2 users. SAMPLE Cell lysates collected from human prostate cancer cell lines PRIMARY ANTIBODY Abcam Ab7747: dilution of 100 microlitres in 15ml TBST + 5% marvel, incubation overnight at 4 degrees. Washes: 1x15 mins and 3x5 mins in TBST. These conditions have worked before. SECONDARY ANTIBODY anti-rabbit, 1:2000 dilution in TBST +5% marvel, incubated for 1 hour at room temp. Washes as before These conditions have worked before. DETECTION METHOD Have tried ECL, ECLPlus and Supersignal. None of these give a band at the correct size. POSITIVE AND NEGATIVE CONTROLS USED Samples that gave a positive result with a different batch of antibody. Cells treated with siRNA against IL8 as negative control ANTIBODY STORAGE CONDITIONS Aliquotted at -20 celcius SAMPLE PREPARATION RIPA with PI tablet + sodium orthovanadate ELB with PI tablet Samples mixed with loading buffer + mercaptoethanol & heated to 95 degrees for 5 minutes. AMOUNT OF PROTEIN LOADED 40 micrograms ELECTROPHORESIS/GEL CONDITIONS 20% reducing gel These conditions have worked before. TRANSFER AND BLOCKING CONDITIONS Transfer for 2 hours in transfer buffer containing 40% methanol. Membrane checked using Ponceau-s. Blocked in TBST +5% marvel for 2 hours. These conditions have worked before. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes ADDITIONAL NOTES I have tried the western 3 times and another user has tried twice. I have tried samples that gave a positive result with a previous batch of antibody. Different sample preparation buffers give no different result. e see a non-specific band arounf 40kDa (have seen this before) but no IL8 band.

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Answer

Thank you for your patience. I am working with the originator of ab7747 to help determine what may be going on with this particular antibody. I would like to confirm, with this batch of ab7747 you never saw an Il-8 band and the only band you saw was at approximately 40 kDa, correct? Did you ever try other positive cells as activated macrophages? Can you give me the lot number that you received - it will be on the vial. Many thanks.

Read More

Answer

The MW of IL-8 are: predicted 11.1 kDa, expressed 6-8 kDa. Please note that IL-8 appears to exist as a dimer in solution. We would recommend 10-15% SDS for acrylamide gel.

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11-14 of 14 Abreviews or Q&A

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