Detects human CCL19/MIP-3 beta in ELISAs and Western blots. In sandwich immunoassays, less than 0.05% cross-reactivity with recombinant mouse (rm) MIP‑3 beta, recombinant human (rh) GRO alpha, rhHCC-4, rhEotaxin, rmMIP-3 alpha, recombinant rat MIP-3 alpha, and rhMIP-3 alpha is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

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

Scientific Data Images for Human CCL19/MIP-3 beta Antibody

Detection of Human CCL19/MIP‑3 beta by Western Blot.

Western blot shows conditioned media from monocyte-derived macrophages untreated (-) or treated (+) with 1 µg/mL LPS and 40 ng/mL Recombinant Human IFN-gamma (Catalog # 285-IF) for 24 hours. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human CCL19/MIP-3 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF361) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CCL19/MIP-3 beta at approximately 12 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Chemotaxis Induced by CCL19/MIP‑3 beta and Neutrali-zation by Human CCL19/MIP‑3 beta Antibody.

Re-combinant Human CCL19/MIP-3 beta (Catalog # 361-MI) chemoattracts the BaF3 mouse pro-B cell line transfected with human CCR7 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 CCL19/MIP-3 beta (50 ng/mL) is neutralized (green line) by increasing concentrations of Human CCL19/MIP-3 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF361). The ND₅₀ is typically 2-10 µg/mL.

Detection of Human CCL19/MIP‑3 beta by Simple Western^TM.

Simple Western lane view shows conditioned media from monocyte-derived macrophages untreated (-) or treated (+) with 1 µg/mL LPS and 40 ng/mL Recombinant Human IFN-gamma (Catalog # 285-IF) for 24 hours. A specific band was detected for CCL19/MIP-3 beta at approximately 8 kDa (as indicated) using 50 µg/mL of Goat Anti-Human CCL19/MIP-3 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF361) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

View 2 more images

Applications for Human CCL19/MIP-3 beta Antibody

Application

Recommended Usage

Simple Western

50 µg/mL
Sample: Conditioned media from monocyte-derived macrophages treated with Recombinant Human IFN‑ gamma
(Catalog # 285-IF)

Western Blot

1 µg/mL
Sample: Recombinant Human CCL19/MIP-3 beta (Catalog # 361-MI)

Neutralization

Measured by its ability to neutralize CCL19/MIP-3 beta-induced chemotaxis in the BaF3 mouse pro-B cell line transfected with human CCR7. The Neutralization Dose (ND₅₀) is typically 2-10 µg/mL in the presence of 50 ng/mL Recombinant Human CCL19/MIP-3 beta.

Human CCL19/MIP-3 beta Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)

Recommended Concentration: 0.2-0.8 µg/mL
Use in combination with these reagents:

Detection Reagent: Human CCL19/MIP‑3 beta Biotinylated Antibody (Catalog # BAF361)
Standard: Recombinant Human CCL19/MIP-3 beta Protein (Catalog # 361-MI)

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 2 using AF361 in the following applications:

Flow Cytometry (1 Review)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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

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: CCL19/MIP-3 beta

MIP-3 beta, also known as ELC (EBI1-Ligand Chemokine), is one of many novel beta chemokines identified through bioinformatics. MIP-3 beta cDNA encodes a 98 amino acid (aa) residue precursor protein with a predicted 21 aa residue signal peptide that is cleaved to form the 77 aa residue mature secreted protein. MIP-3 beta is distantly related to other beta chemokines (20-30% aa sequence identity) and the gene for MIP-3 beta has been mapped to chromosome 9p13 rather than chromosome 17 where the genes for many human beta chemokines are clustered. MIP-3 beta has been shown to be constitutively expressed in various lymphoid tissues (including thymus, lymph nodes, appendix and spleen). The expression of MIP-3 beta is down-regulated by the anti-inflammatory cytokine IL-10. MIP-3 beta has been shown to be a unique functional ligand for CCR7 (previously referred to as the Epstein-Barr virus-induced gene 1 (EBI1) orphan receptor), a chemokine receptor that is expressed in various lymphoid tissues and activated B and T lymphocytes. EBI1 is strongly up-regulated in B cells infected with Epstein-Barr virus and T cells infected with herpesvirus 6 or 7.

Alternate Names

ELC, MIP-3 beta, MIP3 beta

Entrez Gene IDs

6363 (Human); 24047 (Mouse); 362506 (Rat)

Gene Symbol

CCL19

UniProt

Q99731

Additional CCL19/MIP-3 beta Products

Product Documents for Human CCL19/MIP-3 beta Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human CCL19/MIP-3 beta Antibody

For research use only

Citations
Reviews

Human CCL19/MIP-3 beta Antibody

R&D Systems, part of Bio-Techne | Catalog # AF361

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications