Human CCL3/MIP-1 alpha Antibody

Name: Human CCL3/MIP-1 alpha Antibody
Brand: R&D Systems
SKU: AF-270-NA
Rating: 5 (2 reviews)

R&D Systems, Inc. a Bio-Techne Brand

Catalog # AF-270-NA

Citations (12)

(2)

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Key Product Details

Species Reactivity:	Human
Immunogen:	E. coli-derived recombinant human CCL3/MIP-1 alpha Ala27-Ala92 Accession # P10147
Applications:	Block/Neutralize, ELISA Capture, Immunohistochemistry, Western Blot
Label:	Unconjugated
Source:	Polyclonal Goat IgG
Specificity:	Detects human CCL3/MIP‑1 alpha in ELISAs and Western blots. In sandwich immunoassays, less than 0.05% cross-reactivity with recombinant human (rh) MIP-1 beta, rhMIP-1δ, rhMIP-3 alpha, rhMIP-3 beta, and recombinant mouse (rm) MIP-1 alpha is observed. Neutralizes the biological activity of recombinant human CCL3/MIP‑1 alpha. In the chemotaxis assay this antibody will also partially neutralize the biological activity of rhMIP-1 beta at a 20 fold higher IgG concentration. It will not neutralize the biological activity of rmMIP-1 beta or rmMIP-1 alpha.

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Applications for Human CCL3/MIP-1 alpha Antibody

	Recommended Concentration	Sample
Western Blot	0.1 µg/mL	Recombinant Human CCL3/MIP‑1 alpha isoform LD78a (Catalog # 270-LD)
Immunohistochemistry	5-15 µg/mL	See below

Human CCL3/MIP-1 alpha Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

0.2-0.8 µg/mL

Use in combination with:

Detection Reagent: Human CCL3/MIP‑1 alpha Biotinylated Antibody (Catalog # BAF270)

Standard: Recombinant Human CCL3/MIP-1 alpha Protein (Catalog # 270-LD)

Item	Value
Neutralization	Measured by its ability to neutralize CCL3/MIP‑1 alpha -induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CCR5. The Neutralization Dose (ND₅₀) is typically 2-8 µg/mL in the presence of 0.1 µg/mL Recombinant Human CCL3/MIP‑1 alpha isoform LD78a.

Please Note: Optimal dilutions should be determined by each laboratory for each application.

Scientific Data Examples for Human CCL3/MIP-1 alpha Antibody

	Neutralization Chemotaxis Induced by CCL3/MIP‑1 alpha and Neutral-ization by Human CCL3/ MIP‑1 alpha Antibody. Recombinant Human CCL3/MIP-1a (Catalog # 270-LD) 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 Human CCL3/MIP-1a (0.1 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human CCL3/MIP-1a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-270-NA). The ND₅₀ is typically 2-8 µg/mL.
	Immunohistochemistry CCL3/MIP‑1 alpha in Human Tonsil. CCL3/MIP-1a was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human CCL3/MIP-1a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-270-NA) at 10 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 in lymphocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Formulation, Preparation and Storage

Purification	Antigen Affinity-purified
Endotoxin level	<0.10 EU per 1 μg of the antibody by the LAL method.
Reconstitution	Reconstitute at 0.2 mg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
Formulation	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.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & 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.

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Background: CCL3/MIP-1 alpha

The macrophage inflammatory proteins -1 alpha and -1 beta were originally co-purified from medium conditioned by an LPS-stimulated murine macrophage cell line. Human MIP-1 alpha refers to the products of several independently cloned cDNAs, including LD78, pL78, pAT464, and GOS19. These cDNAs all code for the same human protein that is a homologue of the murine MIP-1 alpha. Mature MIP-1 alpha and MIP-1 beta in both human and mouse share approximately 70% homology at the amino acid level. 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. The functional receptor for MIP-1 alpha has been identified as CCR1 and CCR5.

Item	Value
References	Menten, P. et al. (2002) Cytokine Growth Factor Rev. 13:455.
Entrez Gene IDs	6348 (Human); 20302 (Mouse); 25542 (Rat); 448787 (Canine); 102136134 (Cynomolgus Monkey)
Alternate Names	C-C motif chemokine 3, MIP1-(a), AI323804, CCL3, G0S19-1, LD78a, LD78alpha, MIP-1 alpha, MIP-1alpha, MIP1 alpha, MIP1-alpha, MIP1A, PAT 464.1, SCYA3, SIS-beta, chemokine (C-C motif) ligand 3

Citations for Human CCL3/MIP-1 alpha Antibody (12)

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Citations are publications that use Bio-Techne products. Selected citations for Human CCL3/MIP-1 alpha Antibody include:

Species: Human
Sample Types: Whole Cells
Applications: Neutralization

T Kodama et al. (2020), CCL3-CCR5 axis contributes to progression of esophageal squamous cell carcinoma by promoting cell migration and invasion via Akt and ERK pathways Lab. Invest., 0(0).
PMID: 32457351
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry

T Leng et al. (2019), TCR and Inflammatory Signals Tune Human MAIT Cells to Exert Specific Tissue Repair and Effector Functions Cell Rep, 28(12):3077-3091.e5.
PMID: 31533032
Species: Human
Sample Types: Whole Cells
Applications: Neutralization

Walker W et al. (2015), Increased Levels of Macrophage Inflammatory Proteins Result in Resistance to R5-Tropic HIV-1 in a Subset of Elite Controllers. J Virol, 89(10):5502-14.
PMID: 25740989
Species: Human
Sample Types: Cell Culture Supernates
Applications: ELISA Development

Wu W et al. (2009), Resistance of human alveolar macrophages to Bacillus anthracis lethal toxin. J. Immunol., 183(9):5799-806.
PMID: 19812208
Species: Human
Sample Types: Urine
Applications: Antibody Array Development

Hu H et al. (2009), Noninvasive detection of acute and chronic injuries in human renal transplant by elevation of multiple cytokines/chemokines in urine. Transplantation, 87(12):1814-20.
PMID: 19543058
Species: Human
Sample Types: Cell Culture Supernates
Applications: Luminex Development

Biancotto A et al. (2008), Upregulation of human cytomegalovirus by HIV type 1 in human lymphoid tissue ex vivo. AIDS Res. Hum. Retroviruses, 24(3):453-62.
PMID: 18327985
Species: Human
Sample Types: Cell Culture Supernates
Applications: Luminex Development

Biancotto A et al. (2007), Abnormal activation and cytokine spectra in lymph nodes of people chronically infected with HIV-1. Blood, 109(10):4272-9.
PMID: 17289812
Species: Human
Sample Types: Serum
Applications: Luminex Development

Pfleger C et al. (2007), Effect of serum content and diluent selection on assay sensitivity and signal intensity in multiplex bead-based immunoassays. J. Immunol. Methods, 329(1):214-8.
PMID: 17964594
Species: Human
Sample Types: Plasma
Applications: ELISA Development

Qui H et al. (2007), Fluorescence single-molecule counting assays for high-sensitivity detection of cytokines and chemokines. Clin. Chem., 53(11):2010-2.
PMID: 18030696
Species: Human
Sample Types: Cell Culture Supernates
Applications: ELISA Development

Masih-Khan E et al. (2006), MIP-1alpha (CCL3) is a downstream target of FGFR3 and RAS-MAPK signaling in multiple myeloma. Blood, 108(10):3465-71.
PMID: 16849642
Species: Human
Sample Types: Whole Cells
Applications: Neutralization

Townson JR et al. (2002), Gene copy number regulates the production of the human chemokine CCL3-L1. Eur. J. Immunol., 32(10):3016-26.
PMID: 12355456
Species: Human
Sample Types: Whole Cells
Applications: Neutralization

Binninger-Schinzel D et al. (2002), Simian immunodeficiency viruses with defective nef genes show increased susceptibility to the noncytotoxic antiviral activity of CD8+ lymphocytes. , 294(1):209-21.
PMID: 11886279

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Customer Reviews for Human CCL3/MIP-1 alpha Antibody (2)

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Human CCL3/MIP-1 alpha Antibody

Name: Anonymous

Application: ELISA

Sample Tested: Cell culture supernatant

Species: Human

Verified Customer | Posted 09/07/2019
Human CCL3/MIP-1 alpha Antibody

Name: Anonymous

Application: ELISA

Sample Tested: Cell Lysates,RAW 264.7 mouse monocyte/macrophage cell line

Species: Human

Verified Customer | Posted 08/10/2019

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Human CCL3/MIP-1 alpha Antibody

R&D Systems, Inc. a Bio-Techne Brand

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Applications for Human CCL3/MIP-1 alpha Antibody

Human CCL3/MIP-1 alpha Sandwich Immunoassay

Scientific Data Examples for Human CCL3/MIP-1 alpha Antibody

Neutralization

Immunohistochemistry

Formulation, Preparation and Storage

Reconstitution Calculator

Background: CCL3/MIP-1 alpha

Citations for Human CCL3/MIP-1 alpha Antibody (12)

Customer Reviews for Human CCL3/MIP-1 alpha Antibody (2)

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Human CCL3/MIP-1 alpha Antibody

Human CCL3/MIP-1 alpha Antibody

Product Documents for Human CCL3/MIP-1 alpha Antibody

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