CCL3/MIP-1 alpha Antibody [Unconjugated]

Western blot shows lysates of RAW 264.7 mouse monocyte/macrophage cell line untreated (-) or treated (+) with 10 µg/mL LPS for 4 hours. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse CCL3/MIP-1a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-450-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CCL3/MIP-1a at approximately 12 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

CCL3/MIP-1a was detected in perfusion fixed frozen sections of mouse thymus using Goat Anti-Mouse CCL3/MIP-1a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-450-NA) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

CCL3/MIP-1a was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line using Goat Anti-Mouse CCL3/MIP-1a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-450-NA) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Recombinant MouseCCL3/MIP‑1 alpha (Catalog #450-MA) chemoattracts the BaF3 mouse pro‑B cell line transfected with human CCR5 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 Mouse CCL3/MIP‑1 alpha (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse CCL3/MIP‑1 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-450-NA). The ND50 is typically0.05-0.2 µg/mL.

• Reactivity: Mu
• Applications: WB, IHC, B/N, ELISA(Cap), ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

CCL3/MIP-1 alpha Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant mouse CCL3/MIP-1 alpha
  Ala24-Ala92
  Accession # Q5QNW0
• Specificity: Detects mouse CCL3/MIP-1 alpha in ELISAs and Western blots. In sandwich ELISAs, less than 0.03% cross-reactivity with recombinant human (rh) CCL3, recombinant mouse (rm) CCL9/10, and rmCCL4 and less than 0.4% cross-reactivity with rhCCL7.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CCL3
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • ELISA Capture (Matched Antibody Pair) 0.2-0.8 ug/mL
  • ELISA Detection (Matched Antibody Pair) 0.1-0.4 ug/mL
  • ELISA Standard (Matched Pair)
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Neutralization 0.05-0.2 ug/mL
  • Western Blot 1 ug/mL
• Application Notes: ELISA Capture: Mouse CCL3/MIP-1 alpha Antibody (Catalog # AF-450-NA)
  ELISA Detection: Mouse CCL3/MIP-1 alpha Biotinylated Antibody (Catalog # BAF450)
  Standard: Recombinant Mouse CCL3/MIP-1 alpha (Catalog # 450-MA)

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C 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 CCL3/MIP-1 alpha Antibody [Unconjugated]

• C-C motif chemokine 3
• MIP1-(a)
• AI323804
• CCL3
• chemokine (C-C motif) ligand 3
• G0S19-1
• LD78a
• LD78alpha
• MIP1 alpha
• MIP-1 alpha
• MIP1A
• MIP-1alpha
• MIP1-alpha
• PAT 464.1
• SCYA3
• SIS-beta

Background

The macrophage inflammatory proteins 1 alpha and 1 beta , two closely related but distinct proteins, were originally co-purified from medium conditioned by a LPS-stimulated murine macrophage cell line. Mature mouse MIP-1 alpha shares approximately 77% and 70% amino acid identity with human MIP-1 alpha and mouse MIP-1 beta , respectively. MIP‑1 proteins are expressed primarily in T cells, B cells, and monocytes after antigen or mitogen stimulation. The MIP-1 proteins are members of the beta (C-C) subfamily of chemokines.

Both MIP-1 alpha and MIP-1 beta are monocyte chemoattractants in vitro. Additionally, the MIP-1 proteins have been reported to have chemoattractant and adhesive effects on lymphocytes, with MIP-1 alpha and MIP-1 beta preferentially attracting CD8⁺ and CD4⁺ T cells, respectively. MIP-1 alpha has also been shown to attract B cells as well as eosinophils. MIP-1 proteins have been reported to have multiple effects on hematopoietic precursor cells and MIP-1 alpha  has been identified as a stem cell inhibitory factor that can inhibit the proliferation of hematopoietic stem cells in vitro as well as in vivo. In the same assays, MIP-1 beta was reported to be much less active. The functional receptor for MIP-1 alpha has been identified as CCR1 and CCR5.

1. Menten, P. et al. (2002) Cytokine Growth Factor Rev. 13:455.

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 CCL3/MIP-1 alpha Antibody (AF-450-NA)(28)

Publications using AF-450-NA

• Zhang, H;Wang, Z;Li, J;Jia, Y;Li, F; Timing, initiation and function: An in-depth exploration of the interaction network among neutrophil extracellular traps related genes in acute pancreatitis International immunopharmacology 2024-08-12 [PMID: 39137629] (In vivo assay, Mouse)
• Ciechanowska, A;Pawlik, K;Ciapa?a, K;Mika, J; Pharmacological Modulation of the MIP-1 Family and Their Receptors Reduces Neuropathic Pain Symptoms and Influences Morphine Analgesia: Evidence from a Mouse Model Brain sciences 2023-03-29 [PMID: 37190544] (In Vivo, Mouse)
• Van Anh Do-Thi, Sang Min Park, Song Mi Park, Hye Jin Jeong, Geunyoung Cho, Hyun-Jung An et al. IL9 Polarizes Macrophages to M1 and Induces the Infiltration of Antitumor Immune Cells via MIP-1 and CXCR3 Chemokines Cancer Research Communications 1/18/2023 [PMID: 36968220]
• Xi Yuan, Zihan Xiong, Wei Liu, Yue Li, Hongdong Li, Xuemei Zhang et al. Novel Therapeutic Targeting of CCL3-CCR4 Axis Mediated Apoptotic Intesitnal Injury in Necrotizing Enterocolitis Frontiers in Immunology 4/22/2022 [PMID: 35529858]
• Jessica M. Rosin, Nikol Tretiakov, Emily Hanniman, Kiana Hampton, Deborah M. Kurrasch Gestational Bisphenol A Exposure Impacts Embryonic Hypothalamic Microglia Numbers, Ramification, and Phagocytic Cups Frontiers in Neuroscience 2/18/2022 [PMID: 35250464]
• P Bhattachar, N Ismail, A Saxena, S Gannavaram, R Dey, T Oljuskin, A Akue, K Takeda, J Yu, S Karmakar, PK Dagur, JP McCoy, HL Nakhasi Neutrophil-dendritic cell interaction plays an important role in live attenuated Leishmania vaccine induced immunity PloS Neglected Tropical Diseases, 2022-02-22;16(2):e0010224. 2022-02-22 [PMID: 35192633] (In Vivo, Mouse)
• Jorge Luis Galeano Niño, Sophie V Pageon, Szun S Tay, Feyza Colakoglu, Daryan Kempe, Jack Hywood et al. Cytotoxic T cells swarm by homotypic chemokine signalling eLife 10/13/2020 [PMID: 33046212]
• AM Osman, Y Sun, TC Burns, L He, N Kee, N Oliva-Vila, A Alevyzaki, K Zhou, L Louhivuori, P Uhlén, E Hedlund, C Betsholtz, VM Lauschke, J Kele, K Blomgren Radiation Triggers a Dynamic Sequence of Transient Microglial Alterations in Juvenile Brain Cell Rep, 2020-06-02;31(9):107699. 2020-06-02 [PMID: 32492415] (IHC, Mouse)
• Hasan M. Sayeed, Eun‐So Lee, Hae‐Ok Byun, Seonghyang Sohn The role of CCR1 and therapeutic effects of anti‐CCL3 antibody in herpes simplex virus‐induced Behçet's disease mouse model Immunology 11/1/2019 [PMID: 31393598]
• Krisztián Pajer, Tamás Bellák, Heinz Redl, Antal Nógrádi Neuroectodermal Stem Cells Grafted into the Injured Spinal Cord Induce Both Axonal Regeneration and Morphological Restoration via Multiple Mechanisms Journal of Neurotrauma 11/1/2019 [PMID: 31111776]
• B John, C Naczki, C Patel, A Ghoneum, S Qasem, Z Salih, N Said Regulation of the bi-directional cross-talk between ovarian cancer cells and adipocytes by SPARC Oncogene, 2019-02-14;0(0):. 2019-02-14 [PMID: 30765860] (Neutralization, Mouse)
• W Sun, N Meednu, A Rosenberg, J Rangel-Mor, V Wang, J Glanzman, T Owen, X Zhou, H Zhang, BF Boyce, JH Anolik, L Xing B cells inhibit bone formation in rheumatoid arthritis by suppressing osteoblast differentiation Nat Commun, 2018-12-03;9(1):5127. 2018-12-03 [PMID: 30510188] (Neutralization, Mouse)
• VE Thaney, AM O'Neill, MM Hoefer, R Maung, AB Sanchez, M Kaul IFN? Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury Sci Rep, 2017-04-20;7(0):46514. 2017-04-20 [PMID: 28425451] (Neutralization, Mouse, Rat)
• David Blum Hippocampal T cell infiltration promotes neuroinflammation and cognitive decline in a mouse model of tauopathy Brain, 2016-11-05;0(0):. 2016-11-05 [PMID: 27818384] (IHC, Mouse)
• Krisztián Pajer, Georg A. Feichtinger, Gábor Márton, Sonja Sabitzer, Dieter Klein, Heinz Redl et al. Cytokine signaling by grafted neuroectodermal stem cells rescues motoneurons destined to die Experimental Neurology 11/1/2014 [PMID: 24907401]
• Zhang M, Yin L, Zhang K, Sun W, Yang S, Zhang B, Salzman P, Wang W, Liu C, Vidyasagar S, Zhang L, Ju S, Okunieff P, Zhang L Response patterns of cytokines/chemokines in two murine strains after irradiation. Cytokine, 2012-01-25;58(2):169-77. 2012-01-25 [PMID: 22277799] (Luminex Development, Mouse)
• Kim J, Natarajan S, Vaickus LJ, Bouchard JC, Beal D, Cruikshank WW, Remick DG Diesel exhaust particulates exacerbate asthma-like inflammation by increasing CXC chemokines. Am. J. Pathol., 2011-10-01;179(6):2730-9. 2011-10-01 [PMID: 21967814] (ELISA Development, Mouse)
• Eberlein J, Nguyen TT, Victorino F, Golden-Mason L, Rosen HR, Homann D Comprehensive assessment of chemokine expression profiles by flow cytometry. J. Clin. Invest., 2010-02-08;120(3):907-23. 2010-02-08 [PMID: 20197626] (Flow Cytometry, Mouse)
• Diao J, Zhao J, Winter E, Cattral MS Recruitment and differentiation of conventional dendritic cell precursors in tumors. J. Immunol., 2009-12-21;184(3):1261-7. 2009-12-21 [PMID: 20026742] (Neutralization, Mouse)
• Wu W, Mehta H, Chakrabarty K, Booth JL, Duggan ES, Patel KB, Ballard JD, Coggeshall KM, Metcalf JP Resistance of human alveolar macrophages to Bacillus anthracis lethal toxin. J. Immunol., 2009-10-07;183(9):5799-806. 2009-10-07 [PMID: 19812208] (ELISA Development, Mouse)
• Nold-Petry CA, Nold MF, Nielsen JW, Bustamante A, Zepp JA, Storm KA, Hong JW, Kim SH, Dinarello CA Increased cytokine production in IL-18 receptor alpha-deficient cells is associated with dysregulation of suppressors of cytokine signaling (SOCS). J. Biol. Chem., 2009-07-10;0(0):. 2009-07-10 [PMID: 19592492] (ELISA Development, Mouse)
• Wu Y, Li YY, Matsushima K, Baba T, Mukaida N CCL3-CCR5 axis regulates intratumoral accumulation of leukocytes and fibroblasts and promotes angiogenesis in murine lung metastasis process. J. Immunol., 2008-11-01;181(9):6384-93. 2008-11-01 [PMID: 18941229] (IHC-P, Mouse)
• Di Liberto D, Locati M, Caccamo N, Vecchi A, Meraviglia S, Salerno A, Sireci G, Nebuloni M, Caceres N, Cardona PJ, Dieli F, Mantovani A Role of the chemokine decoy receptor D6 in balancing inflammation, immune activation, and antimicrobial resistance in Mycobacterium tuberculosis infection. J. Exp. Med., 2008-08-11;205(9):2075-84. 2008-08-11 [PMID: 18695004] (Neutralization, Mouse)
• Ramos CD, Fernandes KS, Canetti C, Teixeira MM, Silva JS, Cunha FQ Neutrophil recruitment in immunized mice depends on MIP-2 inducing the sequential release of MIP-1alpha, TNF-alpha and LTB(4). Eur. J. Immunol., 2006-08-01;36(8):2025-34. 2006-08-01 [PMID: 16856209] (ELISA Development, Mouse)
• Ellaban E, Bolgos G, Remick D Selective macrophage suppression during sepsis. Cell. Immunol., 2005-02-26;231(1):103-11. 2005-02-26 [PMID: 15919375] (ELISA Development, Mouse)
• Perrin FE, Lacroix S, Aviles-Trigueros M, David S Involvement of monocyte chemoattractant protein-1, macrophage inflammatory protein-1alpha and interleukin-1beta in Wallerian degeneration. Brain, 2005-02-02;128(0):854-66. 2005-02-02 [PMID: 15689362] (Neutralization, Mouse)
• Cardona AE, Gonzalez PA, Teale JM CC chemokines mediate leukocyte trafficking into the central nervous system during murine neurocysticercosis: role of gamma delta T cells in amplification of the host immune response. Infect. Immun., 2003-05-01;71(5):2634-42. 2003-05-01 [PMID: 12704138] (ICC, Mouse)
• Horner AA, Datta SK, Takabayashi K, Belyakov IM, Cinman N, Nguyen MD, Van Uden JH, Berzofsky JA Immunostimulatory DNA-based vaccines elicit multifaceted immune responses against HIV at systemic and mucosal sites. J. Immunol., 2001-08-01;167(3):1584-91. 2001-08-01 [PMID: 11466380] (ELISpot Development, Mouse)

Bioinformatics

• Gene Symbol: CCL3
• Uniprot:
  • Q5QNW0()