Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Functional Assay, Tissue Culture

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all CXCL10/IP-10/CRG-2 Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant mouse CXCL10/IP-10/CRG-2
Ile22-Pro98
Accession # P17515

Specificity

Detects mouse CXCL10/IP-10/CRG-2 in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant human CXCL10 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 Mouse CXCL10/IP-10/CRG-2 Antibody

Detection of Mouse CXCL10/IP‑10/CRG‑2 by Western Blot.

Western blot shows lysates of RAW 264.7 mouse monocyte/macrophage cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse CXCL10/IP-10/CRG-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-466-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for CXCL10/IP-10/CRG-2 at approximately 15 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Chemotaxis Induced by CXCL10/CRG‑2 and Neutral-ization by Mouse CXCL10/ CRG‑2 Antibody.

Recombinant Mouse CXCL10/ CRG-2 (466-CR) chemoattracts the BaF3 mouse pro-B cell line transfected with human CXCR3 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (AR002). Chemotaxis elicited by Recombinant Mouse CXCL10/ CRG-2 (0.5 µg/mL) is neutral-ized (green line) by increasing concentrations of Goat Anti-Mouse CXCL10/CRG-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-466-NA). The ND₅₀ is typically 2.00-25.0 µg/mL.

Detection of Mouse CXCL10/IP-10/CRG-2 by Western Blot

Conditional deletion of astroglial CXCL10. (A) Location of loxP sites in the CXCL10 locus in CXCL10fl/fl mice. (B) Day 17 post-MOG peptide injection (dpi) representative images of immunostaining for CXCL10-Red, GFAP-Green, and DAPI-Blue. (C) Quantification of images for CXCL10 integrated density at 17 dpi (n = 4–7 mice/group). (D and E) qRT/PCR for CXCL10 and CXCL9 in spinal cord of astroglial CXCL10 knockout and littermate control mice at 17 dpi (n = 6 mice/group). (F) Quantification of CXCL10 protein levels in the spinal cord of astroglial CXCL10 knockout and control mice at 14 dpi, by western blot analysis (n = 4 mice/group). (G) Data were normalized to GAPDH, P = 0.046. Vertical bars = SEMs. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24924222), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse CXCL10/IP-10/CRG-2 by Immunocytochemistry/Immunofluorescence

Detection of Mouse CXCL10/IP-10/CRG-2 by Immunohistochemistry

Correlations between neutrophil/mast cell numbers and CXCL10 expression in the fracture hematoma during the early inflammatory phase. Correlation analysis by simple linear regression was performed between the parameters neutrophil numbers, mast cell numbers and local CXCL10 protein expression in all samples. (A) Neutrophil number/CXCL10 correlation. (B) Mast cell number/CXCL10 correlation. (C) Mast cell number/neutrophil number correlation. (D) Immunofluoresence double staining for mast cells (Avidin staining, red) and CXCL10 (green). DNA was counterstained with Hoechst (blue). Scale bar = 50 µm. E) Black box marked the area which is shown in (E). (F) Single fluorescent channels for Avidin (red) and CXCL10 (green). Scale bar = 50 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36761764), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse CXCL10/IP-10/CRG-2 by Immunohistochemistry

Mouse CXCL10 / IP-10 / CRG-2 ELISA Standard Curve

Recombinant Mouse CXCL10/IP-10/CRG-2 (Catalog # 466-CR) was serially diluted and captured by Rat Anti-Mouse CXCL10/IP-10/CRG-2 Monoclonal Antibody (Catalog # MAB466) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Mouse CXCL10/IP-10/CRG-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-466-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

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Applications for Mouse CXCL10/IP-10/CRG-2 Antibody

Application

Recommended Usage

Western Blot

1 µg/mL
Sample: RAW 264.7 mouse monocyte/macrophage cell line

Neutralization

Measured by its ability to neutralize CXCL10/IP‑10/CRG‑2-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CXCR3. The Neutralization Dose (ND₅₀) is typically 2.00-25.0 µg/mL in the presence of 0.5 µg/mL Recombinant Mouse CXCL10/IP‑10/CRG‑2.

Reviewed Applications

Read 3 reviews rated 2.7 using AF-466-NA in the following applications:

ELISA (1 Review)
Immunocytochemistry/Immunofluorescence (1 Review)
Immunohistochemistry-Frozen (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. See Certificate of Analysis for details.
*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: CXCL10/IP-10/CRG-2

The gene for CRG-2, a mouse homolog of human IP-10, was originally identified as an immediate early gene induced in response to macrophage activation. It has since been shown that CRG-2 mRNA is induced by alpha/ beta/ gamma-interferons and by lipopolysaccharide in macrophages, astrocytes and microglia. Human IP-10 was also shown to be expressed in activated T-lymphocytes, splenocytes, keratinocytes, osteoblasts, astrocytes, and smooth muscle cells. Mouse CRG-2 cDNA encodes a 98 amino acid (aa) residue precursor protein with a 21 aa residue signal peptide that is cleaved to form the 77 aa residue secreted mature protein. Mature CRG-2 shares approximately 67% amino acid sequence identity with human IP-10. The amino acid sequence of CRG-2 identified the protein as a member of the chemokine alpha subfamily that lacks the ELR domain. CRG-2 has been shown to be a chemoattractant for activated T-lymphocytes. Recently, human IP-10 has also been reported to be a potent inhibitor of angiogenesis and to display a potent thymus-dependent anti-tumor effect. A chemokine receptor specific for IP-10 and MIG (CXCR3) has been cloned and shown to be highly expressed in IL-2-activated T-lymphocytes.

References

Loetscher, M. et al. (1996) J. Exp. Med. 184:963.
Vanguri, P. (1996) J. Neuroimmunol. 56:35.
Sgadari, C. et al. (1996) Blood 87:3877.

Alternate Names

CRG-2, CRG2, IP-10

Entrez Gene IDs

3627 (Human); 15945 (Mouse)

Gene Symbol

CXCL10

UniProt

P17515

Additional CXCL10/IP-10/CRG-2 Products

Product Documents for Mouse CXCL10/IP-10/CRG-2 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Mouse CXCL10/IP-10/CRG-2 Antibody

For research use only

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