CXCL8/IL-8 Antibody [Unconjugated]

Western blot shows lysates and conditioned media of THP‑1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA and 10 ug/mL LPS for 24 hours and 3 hours. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human IL-8/CXCL8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-208-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for IL-8/CXCL8 at approximately 10 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Simple Western lane view shows PBMC conditioned media untreated (-) or treated (+) with 1 μg/mL LPS for 24 hrs, loaded at 0.2 mg/mL. A specific band was detected for IL-8/CXCL8 at approximately 8 kDa (as indicated) using 25 µg/mL of Goat Anti-Human IL-8/CXCL8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-208-NA) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF019). This experiment was conducted under reducing conditions and using the 2-40 kDa separation system.

Simple Western lane view shows lysates of THP‑1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA for 24 hrs and 10ug/mL LPS for 3 hrs, loaded at 0.2 mg/mL. A specific band was detected for IL-8/CXCL8 at approximately 8 kDa (as indicated) using 25 µg/mL of Goat Anti-Human IL-8/CXCL8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-208-NA) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF019). This experiment was conducted under reducing conditions and using the 2-40 kDa separation system.

Recombinant Human IL-8/CXCL8 (Catalog # 208-IL) chemoattracts the BaF3 mouse pro‑B cell line transfected with human CXCR2 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Human IL-8/CXCL8 (20 ng/mL) is neutralized (green line) by increasing concentrations of Human IL-8/CXCL8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-208-NA). The ND50 is typically0.1‑0.5 µg/mL.

IL-8/CXCL8 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with PMA and ionomycin using 10 µg/mL Human IL-8/CXCL8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-208-NA) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

IL-8/CXCL8 was detected in immersion fixed paraffin-embedded sections of human tonsil (surgically removed due to severe EBV-induced mononucleosis) using Human IL-8/CXCL8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF‑208‑NA) overnight at 4 °C. Tissue was stained (brown) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

• Reactivity: Hu
• Applications: WB, Simple Western, IHC, B/N, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

CXCL8/IL-8 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human IL-8/CXCL8
  Ser28-Ser99
  Accession # P10145
• Specificity: Detects human IL-8/CXCL8 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: IL8
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Neutralization 0.1-0.5 ug/mL
  • Simple Western 25 ug/mL
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  12 months from date of receipt, -20 to -70 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CXCL8/IL-8 Antibody [Unconjugated]

• 3-10C
• AMCF-I
• C-X-C motif chemokine 8
• CXCL8
• CXCL8SCYB8
• Emoctakin
• GCP1
• GCP-1TSG-1
• IL8
• IL-8
• interleukin 8
• K60
• LAI
• LECT
• LUCT
• LYNAP
• MDNCF
• MDNCFb-ENAP
• member 8
• MONAP
• MONAPGCP1
• NAF
• NAP1
• NAP-1NAP1
• NCF
• Neutrophil-activating protein 1
• Protein 3-10C
• T cell chemotactic factor
• T-cell chemotactic factor
• TCF
• TSG1

Background

CXCL8 was originally discovered and purified independently by a number of laboratories as a neutrophil chemotactic and activating factor. It was also referred to as neutrophil chemotactic factor (NCF), neutrophil activating protein (NAP), monocyte-derived neutrophil chemotactic factor (MDNCF), T-lymphocyte chemotactic factor (TCF), granulocyte chemotactic protein (GCP) and leukocyte adhesion inhibitor (LAI). Many cell types, including monocyte/macrophages, T cells, neutrophils, fibroblasts, endothelial cells, keratinocytes, hepatocytes, chondrocytes, and various tumor cell lines, can produce CXCL8 in response to a wide variety of pro-inflammatory stimuli such as exposure to IL-1, TNF, LPS, and viruses. CXCL8 is a member of the alpha (CXC) subfamily of chemokines, which also includes platelet factor 4, GRO, IP-10, etc. CXCL8 is a potent chemoattractant for neutrophils. In addition, CXCL8 also has a wide range of other pro-inflammatory effects. CXCL8 causes degranulation of neutrophil specific granules and azurophilic granules. CXCL8 induces expression of the cell adhesion molecules CD11/CD18 and enhances the adherence of neutrophils to endothelial cells and sub-endothelial matrix proteins. Besides neutrophils, CXCL8 is also chemotactic for basophils, T cells and eosinophils. CXCL8 has been reported to be a co-mitogen for keratinocytes and was also shown to be an autocrine growth factor for melanoma cells. CXCL8 was also reported to be angiogenic both in vivo and in vitro.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CXCL8/IL-8 Antibody (AF-208-NA)(25)

Publications using AF-208-NA

• Franco, YA;de Moraes, MO;Carvalho, LAC;Dohle, W;da Silva, RO;Noma, IHY;Lima, K;Potter, BVL;Machado-Neto, JA;Maria-Engler, SS; 2-Methoxyestradiol-3,17-O,O-bis-sulfamate (STX140) Inhibits Proliferation and Invasion via Senescence Pathway Induction in Human BRAFi-Resistant Melanoma Cells International journal of molecular sciences 2023-07-11 [PMID: 37511073] (Human)
• Zhi-yuan Zhang, Cui-yu Ju, Liu-zheng Wu, Han Yan, Wen-bin Hong, Hang-zi Chen et al. Therapeutic potency of compound RMY-205 for pulmonary fibrosis induced by SARS-CoV-2 nucleocapsid protein Cell Chemical Biology 3/16/2023 [PMID: 36889311]
• H Xiong, J Ye, K Xie, W Hu, N Xu, H Yang Exosomal IL-8 derived from Lung Cancer and Colon Cancer cells induced adipocyte atrophy via NF-kappaB signaling pathway Lipids in Health and Disease, 2022-12-29;21(1):147. 2022-12-29 [PMID: 36581870] (Western Blot, Mouse)
• Haiyan Piao, Lingfeng Fu, Yuxin Wang, Yang Liu, Yue Wang, Xiangyu Meng et al. A positive feedback loop between gastric cancer cells and tumor-associated macrophage induces malignancy progression Journal of Experimental & Clinical Cancer Research 5/14/2022 [PMID: 35562774]
• Ryan Wallis, Natasa Josipovic, Hannah Mizen, Arturo Robles‐Tenorio, Eleanor J. Tyler, Argyris Papantonis et al. Isolation methodology is essential to the evaluation of the extracellular vesicle component of the senescence‐associated secretory phenotype Journal of Extracellular Vesicles 2/18/2021 [PMID: 33659050]
• Youyuan Deng, Zhijie Ning, Zhiya Hu, Qianle Yu, Bin He, Guohuang Hu High interleukin‑8 and/or extracellular signal‑regulated kinase 2 expression predicts poor prognosis in patients with hepatocellular carcinoma Oncology Letters 9/23/2019 [PMID: 31612032]
• Youyuan Deng, Zhijie Ning, Zhiya Hu, Qianle Yu, Bin He, Guohuang Hu High interleukin‑8 and/or extracellular signal‑regulated kinase 2 expression predicts poor prognosis in patients with hepatocellular carcinoma Oncology Letters 2019-09-23 [PMID: 31612032] (Immunohistochemistry, Human)
• Y Chen, M Zhong, G Yuan, H Peng Interleukin-17 induces angiogenesis in�vitro via CXCL8 and CCL2 in retinal pigment epithelium Mol Med Rep, 2018-01-18;0(0):. 1/18/2018 [PMID: 29363728]
• P Ortiz-Mont, A Londoño-Va, JP Vernot Senescence-associated IL-6 and IL-8 cytokines induce a self- and cross-reinforced senescence/inflammatory milieu strengthening tumorigenic capabilities in the MCF-7 breast cancer cell line Cell Commun. Signal, 2017-05-04;15(1):17. 2017-05-04 [PMID: 28472950] (Neutralization, Human)
• LQ Siew, SY Wu, S Ying, CJ Corrigan Cigarette smoking increases bronchial mucosal IL-17A expression in asthmatics, which acts in concert with environmental aeroallergens to engender neutrophilic inflammation Clin. Exp. Allergy, 2017-03-13;0(0):. 2017-03-13 [PMID: 28211191] (IHC, Human)
• SH Ahn, H Park, YH Ahn, S Kim, MS Cho, JL Kang, YH Choi Necrotic cells influence migration and invasion of glioblastoma via NF-?B/AP-1-mediated IL-8 regulation Sci Rep, 2016-04-14;6(0):24552. 2016-04-14 [PMID: 27076368] (IHC-P, Human)
• Maria Hårdstedt, Susanne Lindblom, Alex Karlsson-Parra, Bo Nilsson, Olle Korsgren Characterization of Innate Immunity in an Extended Whole Blood Model of Human Islet Allotransplantation Cell Transplantation 3/1/2016 [PMID: 26084381]
• Maria Hårdstedt, Susanne Lindblom, Alex Karlsson-Parra, Bo Nilsson, Olle Korsgren Characterization of Innate Immunity in an Extended Whole Blood Model of Human Islet Allotransplantation Cell Transplantation 2016-03-01 [PMID: 26084381] (ELISA Detection, Human)
• Mizuki Yamamoto, Yuu Taguchi, Taku Ito-Kureha, Kentaro Semba, Noritaka Yamaguchi, Jun-ichiro Inoue NF-kappa B non-cell-autonomously regulates cancer stem cell populations in the basal-like breast cancer subtype Nature Communications 8/12/2013 [PMID: 23934482]
• Nilka Luisa Díaz, Olga Zerpa, Félix Jacobo Tapia Chemokines and chemokine receptors expression in the lesions of patients with American cutaneous leishmaniasis Mem. Inst. Oswaldo Cruz 6/1/2013 [PMID: 23827991]
• Castriconi R, Dondero A, Bellora F, Moretta L, Castellano A, Locatelli F, Corrias M, Moretta A, Bottino C Neuroblastoma-derived TGF-beta1 modulates the chemokine receptor repertoire of human resting NK cells. J Immunol, 2013-04-10;190(10):5321-8. 2013-04-10 [PMID: 23576682] (Neutralization, Human)
• Inho Choi, Yong Suk Lee, Hee Kyoung Chung, Dongwon Choi, Tatiana Ecoiffier, Ha Neul Lee et al. Interleukin-8 Can Reduce Post-Surgical Lymphedema Formation by Promoting Lymphatic Vessel Regeneration Angiogenesis 1/1/2013 [PMID: 22945845]
• Jhen-Wei Ruan, Yi-Chu Liao, Ingrid Lua, Ming-Hsun Li, Chih-Yi Hsu, Ji-Hshiung Chen Human pituitary tumor-transforming gene 1 overexpression reinforces oncogene-induced senescence through CXCR2/p21 signaling in breast cancer cells Breast Cancer Research 8/1/2012 [PMID: 22789011]
• Heather Kalish, Terry M. Phillips Assessment of chemokine profiles in human skin biopsies by an immunoaffinity capillary electrophoresis chip Methods 2/1/2012 [PMID: 22197729]
• Hardison MT, Galin FS, Calderon CE, Djekic UV, Parker SB, Wille KM, Jackson PL, Oster RA, Young KR, Blalock JE, Gaggar A The presence of a matrix-derived neutrophil chemoattractant in bronchiolitis obliterans syndrome after lung transplantation. J. Immunol., 2009-04-01;182(7):4423-31. 2009-04-01 [PMID: 19299743] (Neutralization, Human)
• Bendrik C, Dabrosin C Estradiol increases IL-8 secretion of normal human breast tissue and breast cancer in vivo. J. Immunol., 2009-01-01;182(1):371-8. 2009-01-01 [PMID: 19109168] (IHC-P, Human)
• Watanabe H, Kawaguchi M, Fujishima S, Ogura M, Matsukura S, Takeuchi H, Ohba M, Sueki H, Kokubu F, Hizawa N, Adachi M, Huang SK, Iijima M Functional characterization of IL-17F as a selective neutrophil attractant in psoriasis. J. Invest. Dermatol., 2008-10-02;129(3):650-6. 2008-10-02 [PMID: 18830271] (In Vivo, Mouse)
• Shahbakhti H, Watson RE, Azurdia RM, Ferreira CZ, Garmyn M, Rhodes LE Influence of eicosapentaenoic acid, an omega-3 fatty acid, on ultraviolet-B generation of prostaglandin-E2 and proinflammatory cytokines interleukin-1 beta, tumor necrosis factor-alpha, interleukin-6 and interleukin-8 in human skin in vivo. Photochem. Photobiol., 2004-09-01;80(2):231-5. 2004-09-01 [PMID: 15362934] (IHC, Human)
• Lukashevich IS, Tikhonov I, Rodas JD, Zapata JC, Yang Y, Djavani M, Salvato MS Arenavirus-mediated liver pathology: acute lymphocytic choriomeningitis virus infection of rhesus macaques is characterized by high-level interleukin-6 expression and hepatocyte proliferation. J. Virol., 2003-02-01;77(3):1727-37. 2003-02-01 [PMID: 12525606] (Neutralization, Primate - Macaca mulatta (Rhesus Macaque))
• Keane MP, Donnelly SC, Belperio JA, Goodman RB, Dy M, Burdick MD, Fishbein MC, Strieter RM Imbalance in the expression of CXC chemokines correlates with bronchoalveolar lavage fluid angiogenic activity and procollagen levels in acute respiratory distress syndrome. J. Immunol., 2002-12-01;169(11):6515-21. 2002-12-01 [PMID: 12444162] (ELISA Development, IHC-P, Human)

Reviews for CXCL8/IL-8 Antibody (AF-208-NA) (4)

Average Rating: 4.5

(Based on 4 reviews)

We have 4 reviews tested in 2 species: Human, Other.

reviewed by: Verified Customer WB Human 01/06/2015

Summary

• Application: Western Blot
• Sample Tested: See PMID 23150618
• Species: Human

reviewed by: Verified Customer WB Other 01/06/2015

Summary

• Application: Western Blot
• Sample Tested: See PMID 19552626
• Species: Other

reviewed by: Verified Customer IF Human 01/06/2015

Summary

• Application: Immunofluorescence
• Sample Tested: See PMID 19258324
• Species: Human

reviewed by: Verified Customer ELISA Human 01/06/2015

Summary

• Application: ELISA
• Sample Tested: See PMID 11744674
• Species: Human

Bioinformatics

• Gene Symbol: IL8
• Uniprot:
  • P10145()