CCL22, also named stimulated T cell chemotactic protein (STCP-1) and MDC, is a CC chemokine initially isolated from clones of monocyte-derived macrophages. Human CCL22 cDNA encodes a precursor protein of 93 amino acid residues with a 24 amino acid residue predicted signal peptide that is cleaved to yield a 69 amino acid residue mature 8 kDa protein. At the amino acid sequence level, CCL22 shows less than 35% identity to other CC chemokine family members. Human CCL22 is expressed in dendritic cells, macrophages and activated monocytes. In addition, CCL22 expression is also detected in the tissues of thymus, lymph node and appendix. The gene for human CCL22 has been mapped to chromosome 16 rather than chromosome 17 where the genes for many human CC chemokines are clustered. Recombinant or chemically synthesized mature CCL22 has been shown to induce chemotaxis or Ca2+ mobilization in dendritic cells, IL-2 activated NK cells, and activated T lymphocytes. A CD8+ T lymphocyte-derived secreted soluble activity that suppresses infection by primary non-syncytium-inducing and syncytium-inducing HIV-1 isolates and the T cell line-adapted isolate HIV-1IIIB, has been identified as CCL22. Based on amino-terminal sequence analysis, the major CD8+ T lymphocyte-derived CCL22 protein yielded an amino-terminal sequence of YGANM, which is two amino acid residues shorter than the predicted mature CCL22. The difference in potency between the two mature CCL22 isoforms has not been determined.
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human
Cited:
Human, Guinea Pig, Transgenic Mouse, Xenograft
Applications
Validated:
Western Blot, ELISA Capture (Matched Antibody Pair), Neutralization
Cited:
Immunohistochemistry-Paraffin, Western Blot, Neutralization, Flow Cytometry, Bioassay, ELISA Capture, ELISA Development
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG2B Clone # 57226
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Product Specifications
Immunogen
E. coli-derived recombinant human CCL22/MDC
Gly25-Gln93
Accession # O00626.1
Gly25-Gln93
Accession # O00626.1
Specificity
Detects human CCL22/MDC in ELISAs and Western blots. In ELISAs, no cross-reactivity with recombinant mouse CCL17, recombinant human (rh) CCL17, or rhCCL21 is observed.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG2B
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Scientific Data Images for Human CCL22/MDC Antibody
Chemotaxis Induced by CCL22/MDC and Neutralization by Human CCL22/MDC Antibody.
Recombinant Human CCL22/MDC (Catalog # 336-MD) chemoattracts the BaF3 mouse pro-B cell line transfected with human CCR4 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 CCL22/MDC (10 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human CCL22/MDC Monoclonal Antibody (Catalog # MAB336). The ND50 is typically 0.6-3.0 µg/mL.
Detection of Human CCL22/MDC by Western Blot
MiR‐23a inhibitor reversed p65 inhibition effects on HBV‐positive xenograft tumor growth. A, Xenografted tumors were harvested after different treatments. B and C, p65 inhibitor treatment significantly retarded tumor growth and miR‐23a inhibitor promoted tumor growth as indicated by the tumor weight and size. The therapeutic effects of p65 inhibitor were abolished by cotreatment with miR‐23a inhibitor. D, CCL22 protein levels from each group were detected by western blotting. E, Statistical results in D. p65 inhibitor led to reduction of CCL22 protein level but miR‐23a inhibitor increased CCL22 level in tumor tissues. F, p65 inhibitor led to reduction of CCL22 mRNA level but miR‐23a inhibitor increased CCL22 mRNA level in tumor tissues. MiR‐23a inhibitor reversed p65 inhibition effects on CCL22 level in tumor tissues. Error bars represented mean ± SD. *P < .05 and **P < .01 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human CCL22/MDC by Western Blot
MiR‐23a, p‐p65, p65, and CCL22 levels were dysregulated in HCC cell lines. A, RT‐qPCR revealed that miR‐23a expression was lowest in the HBV‐positive HepG2.2.15 cell line. B, Highest mRNA level of CCL22 was observed in HepG2.2.15. C, Protein levels of p‐p65, p65, and CCL22 in three cell lines were determined by Western blotting. D, The gray scale analysis of p‐p65, p65, CCL22 and ratio of p‐p65/p65 in three cell lines. Expression of CCL22, p‐p65, and p65 were higher in HBV+ HepG2.2.15 cells than their parental HBV‐ HepG2 cells and normal WRL68 cells. Error bars represented mean ± SD. **P < .01 and *P < .05 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human CCL22/MDC by Western Blot
MiR‐23a inhibited Tregs migration by directly targeting CCL22. A, CCL22 3′UTR contains a putative miR‐23a‐binding site. B, MiR‐23a mimics significantly reduced the luciferase activity tagged with wide type of CCL22 3′UTR. Mutagenesis in the miR‐23a‐binding site abolished the inhibitory actions. C, Transfection of miR‐23a mimics efficiently increased miR‐23a level in both cell lines. D, The effects of miR‐23a overexpression on p‐p65, p65, and CCL22 protein levels in HepG2 and HepG2.2.15 cells were assessed by western blotting. E, The gray scale analysis of p‐p65, p65, CCL22 after miR‐23a overexpression. MiR‐23a mimics transfection did not alter the expression of p65 and its phosphorylation but CCL22 was significantly reduced by miR‐23a mimics. F, MiR‐23a mimics significantly reduced CCL22 mRNA level. G, Overexpression of miR‐23a was sufficient to attenuate Tregs transmigration in both cell lines. Error bars represented mean ± SD. *P < .05, **P < .01, and ***P < .001 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human CCL22/MDC by Western Blot
Lower miR‐23a expression was correlated with higher level of CCL22 expression and intratumoral Treg recruitment in HBV‐positive HCC. A, HBV infection was associated with patient survival rate of HCC. Patients carrying HBV (n = 30) had significantly poorer prognosis than HBV‐ patients (n = 30). B, HBV‐ tissues (n = 30) and HBV+ tissues (n = 30) expressed significantly lower level of miR‐23a. C, HBV− tissues and HBV+ tissues expressed significantly higher mRNA level of CCL22. D, HBV‐ tissues and HBV+ tissues expressed significantly higher mRNA level of Foxp3. E, The protein levels of CCL22, Foxp3, p‐p65, and p65 in normal control, HBV‐ and HBV+ tumor tissues were evaluated by western blotting. F, The gray scale analysis of CCL22, Foxp3, p‐p65, and p65 in normal control, HBV− and HBV+ tumor tissues. In ascending order: normal < HBV− < HBV+. G, Foxp3 signals (red) were colocalized with CD4 (green) signals in tissues. The ratios of Foxp3+CD4+ cells were gradually increased from normal, HBV− HCC to HBV+ HCC tissues. H. MiR‐23a level was inversely correlated with CCL22 expression in HCC tissues, but not in normal control. I. MiR‐23a level was inversely correlated with Foxp3 expression in HCC tissues, but not in normal control. Error bars represented mean ± SD. **P < .01 and *P < .05. HBV+, HCC tissues with HBV infection. HBV−, HCC tissues without HBV infection. Normal, normal liver samples Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human CCL22/MDC by Western Blot
MiR‐23a inhibitor reversed p65 inhibition effects on CCL22 and Tregs recruitment. A, MiR‐23a inhibitor reversed p65 inhibition effects on miR‐23a level. B, The effects of miR‐23a inhibition and p65 inhibitor treatment on p‐p65, p65, and CCL22 protein levels in HepG2 and HepG2.2.15 cells were assessed by western blotting. C, The gray scale analysis of p‐p65, p65, CCL22. MiR‐23a inhibitor reversed p65 inhibition effects on CCL22 protein level but had no effects on p‐p65 and p65. D, MiR‐23a inhibitor reversed p65 inhibition effects on CCL22 mRNA level. E, MiR‐23a inhibitor reversed p65 inhibition effects on Tregs recruitment. F, CCL22 neutralizing antibody reversed miR‐23a inhibitor‐mediated Tregs migration. Error bars represented mean ± SD. *P < .05, **P < .01, and ***P < .001 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human CCL22/MDC by Western Blot
p65 promoted Tregs recruitment via directly modulating miR‐23a. A, A p65‐binding site could be found in miR‐23a promoter region. B, Overexpression of p65 significantly reduced the luciferase activity driven by miR‐23a promoter. Mutagenesis in the p65‐binding site released such repression. C, ChIP assay confirmed that p65 could bind to the promoter region of miR‐23a. D, p65 inhibition upregulated miR‐23a level. E, p65 inhibitor significantly reduced the p‐p65, p65, and CCL22 protein levels in HepG2 and HepG2.2.15 cells. F, The gray scale analysis of p‐p65, p65, CCL22 after p65 inhibitor treatment. G, p65 inhibition dampened CCL22 mRNA level. H, CD4+CD25+ Foxp3+ human Tregs were identified. I, HBV+ HepG2.2.15 cells caused more Tregs to migrate through transwell membrane than HBV− HpeG2 cells. Pretreatment with p65 inhibitor attenuated Tregs transmigration in both cell lines. Error bars represented mean ± SD. *P < .05, **P < .01, and ***P < .001 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Human CCL22/MDC Antibody
Application
Recommended Usage
Western Blot
1 µg/mL
Sample: Recombinant Human CCL22/MDC (Catalog # 336-MD)
Sample: Recombinant Human CCL22/MDC (Catalog # 336-MD)
Neutralization
Measured by its ability to neutralize CCL22/MDC-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CCR4. The Neutralization Dose (ND50) is typically 0.6-3.0 µg/mL in the presence of 10 ng/mL Recombinant Human CCL22/MDC.
Human CCL22/MDC Sandwich Immunoassay
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Reviewed Applications
Read 2 reviews rated 4 using MAB336 in the following applications:
Formulation, Preparation, and Storage
Purification
Protein A or G purified from ascites
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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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.
Calculators
Background: CCL22/MDC
References
- Godiska, R. et al. (1997) J. Exp. Med. 185:1595.
- Chang, M-S. et al. (1997) J. Biol. Chem. 272:25229.
- Pal, R. et al. (1997) Science 278:5338.
Alternate Names
ABCD-1, MDC, STCP-1
Gene Symbol
CCL22
UniProt
Additional CCL22/MDC Products
Product Documents for Human CCL22/MDC Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human CCL22/MDC Antibody
For research use only
Citations for Human CCL22/MDC Antibody
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Customer Reviews for Human CCL22/MDC Antibody (2)
4 out of 5
2 Customer Ratings
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Application: Western BlotSample Tested: T cellsSpecies: HumanVerified Customer | Posted 11/08/2021
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Application: ELISASample Tested: Serum and PlasmaSpecies: HumanVerified Customer | Posted 06/09/2020We used this antibody in an in-house ELISA along with pAb (AF336) and protein (336-MD-025) to detect CCL22 in human serum and plasma. This combination could detect CCL18 in human serum and plasma.
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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Associated Pathways
Th2 Differentiation Pathway
