Human CCL14a/HCC-1 Antibody

Catalog #: AF-324-PB Datasheet

Catalog #	Availability	Size / Price	Qty
AF-324-PB
AF-324-SP

Chemotaxis Induced by CCL14a/HCC‑1 and Neutral-ization by Human CCL14a/HCC‑1 Antibody.

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Human CCL14a/HCC-1 Antibody Summary

Species Reactivity

Human

Specificity

Detects human CCL14a/HCC‑1 in direct ELISAs and Western blots. In direct ELISAs and Western blots (non-reducing conditions), less than 5% cross-reactivity with recombinant human (rh) MCP-2, rhMCP-3, recombinant mouse (rm) C10, and rmMIP-1 beta is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human CCL14a/HCC‑1
Thr20-Asn93
Accession # Q16627

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.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Human CCL14a/HCC‑1 aa 20-93 (Catalog # 324-HC)

Neutralization

Measured by its ability to neutralize CCL14a/HCC‑1-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CCR1. The Neutralization Dose (ND₅₀) is typically 0.15-0.75 µg/mL in the presence of 10 ng/mL Recombinant Human CCL14a/HCC‑1 aa 28‑93.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Neutralization

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Chemotaxis Induced by CCL14a/HCC‑1 and Neutral-ization by Human CCL14a/HCC‑1 Antibody. Recombinant Human CCL14a/HCC-1 (Catalog # 1578-HC) chemoattracts the BaF3 mouse pro-B cell line transfected with human CCR1 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 CCL14a/HCC-1 (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human CCL14a/ HCC-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-324-PB). The ND₅₀ is typically 0.15-0.75 µg/mL.

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Background: CCL14a/HCC-1

HCC-1 (Hemofiltrate CC Chemokine-1) was originally isolated from the hemofiltrate of human patients with chronic renal failure (1). It belongs to the CC chemokine superfamily and has been designated CCL14a. HCC-1/CCL14a cDNA encodes a 93 amino acid (aa) residue precursor with a 19 aa signal peptide that is cleaved to form the 74 aa secreted protein (aa 20‑93). By alternative splicing, a second longer isoform named HCC-3/CCL14b, which includes sequences from exon 3, also exists (2). HCC-1/CCL14a is expressed constitutively in various normal tissues including spleen, liver, muscle, gut and bone marrow. It circulates at nanomolar concentrations in human plasma. Different post-translationally modified HCC-1/CCL14a, including O-glycosylated and N-terminally truncated variants of HCC‑1/CCL14a, have been identified (3, 4). Whereas the 74 aa peptide is a weak CCR1 agonist, the proteolytically processed, truncated HCC‑1/CCL14a (aa 28‑93) is a highly potent agonist of CCR1, CCR5 and to a lesser extent, CCR3. HCC-1/CCL14a (aa 28‑93) promotes chemotaxis of T lymphocytes, monocytes and eosinophils, and inhibits infection of M-tropic human immunodeficiency virus type 1. Activation of the HCC-1/CCL14a precursor to active peptide is mediated by the urokinase type plasminogen activator or plasmin (5).

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