BD Pharmingen™

Technical Data Sheet

Purified Mouse Anti-Human RON (CD136)Product Information

565691Material Number:

MSP Receptor; MSP-R; MST1R; p185-Ron; PTK8 protein tyrosine kinase 8; PTK8Alternate Name:

Size: 0.1 mg

Concentration: 0.5 mg/ml

Clone: Zt/g4

Immunogen: Human RON Transfected Cell Line

Isotype: Mouse IgG1, κ

QC Testing: HumanReactivity:

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

DescriptionThe Zt/g4 monoclonal antibody specifically binds to RON, which is also known as CD136, Macrophage-stimulating protein receptor

(MSP-Receptor, MSP-R), Macrophage stimulating 1 receptor (c-met-related tyrosine kinase) (MST1R), or Protein-tyrosine kinase 8 (PTK8).

RON is a type I transmembrane glycoprotein that belongs to the receptor tyrosine kinase family. It is expressed on cells as a disulfide-linked

heterodimer. RON is expressed on macrophages, keratinocytes, erythroblasts and epithelial cells from the skin, lung, kidney, liver, intestine,

colon, and bone marrow. RON reportedly regulates many physiological processes including the survival, migration, differentiation, and

effector functions of a variety of cell types including epithelial cells, and macrophages. RON binds Macrophage stimulating protein (MSP).

Overexpression of RON may contribute to the malignant transformation of epithelial cells.

Analysis of human RON (CD136) expression.

Left Panel - Flow cytometric analysis of RON (CD136) expression on human SW480 cells. Cells from the human SW480 (Colorectal

adenocarcinoma, ATCC CCL-228) cell line were stained with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 554121; dashed line

histogram) or Purified Mouse Anti-Human RON (CD136) antibody (Cat. No. 565691; solid line histogram) followed by staining with PE Goat

Anti-Mouse Ig (Cat. No. 550589). The fluorescence histogram showing the expression RON (CD136) [Ig isotype control staining] was

derived from gated events with the forward and side light-scatter characteristics of viable SW480 cells. Flow cytometric analysis was

performed using a BD™ LSR II Flow Cytometer System.

Right Panel - Immunohistochemical staining of human small intestine. After fixation of small intestine with acetone, sections were stained

with either Purified Mouse IgG1, κ Isotype Control (Cat. No.550878; Left Image) or Purified Mouse Anti-Human Ron (CD136) (Cat. No.

565691; Right Image). A three-step staining procedure that employs a Biotin Goat Anti-Mouse Immunoglobulin (Cat. No. 550337),

Streptavidin-Horseradish Peroxidase (HRP) (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary

staining reagents. Original magnification: 40×.

Preparation and StorageStore undiluted at 4°C.

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Application Notes

Application

Flow cytometry Routinely Tested

Immunohistochemistry Tested During Development

565691 Rev. 1 Page 1 of 2

Suggested Companion Products

Catalog Number Name CloneSize

554656 Stain Buffer (FBS) 500 mL (none)

554657 Stain Buffer (BSA) 500 mL (none)

554121 Purified Mouse IgG1 κ Isotype Control 0.1 mg MOPC-21

550589 PE Goat Anti-Mouse Ig (Multiple Adsorption) 0.2 mg Polyclonal

550878 Purified Mouse IgG1 κ Isotype Control 1 mL MOPC-31C

550337 Biotin Goat Anti-Mouse Ig (Multiple Adsorption) 0.25 mg Polyclonal

550946 Streptavidin HRP 50 mL (none)

550880 DAB Substrate Kit 500 Tests (none)

565692 PE Mouse Anti-Human RON (CD136) 50 µg Zt/g4

Product NoticesSince applications vary, each investigator should titrate the reagent to obtain optimal results. 1.

Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols. 2.

Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before

discarding to avoid accumulation of potentially explosive deposits in plumbing.

3.

Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not

be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or

dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the

NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.

4.

An isotype control should be used at the same concentration as the antibody of interest. 5.

ReferencesFeng L, Yao HP, Wang W, et al. Efficacy of anti-RON antibody Zt/g4-drug maytansinoid conjugation (Anti-RON ADC) as a novel therapeutics for targeted

colorectal cancer therapy. Clin Cancer Res. 2014; 20(23):6045-6058. (Clone-specific: Bioassay, Flow cytometry, Fluorescence microscopy, Functional assay,

Immunofluorescence, In vivo exacerbation)

Li Z, Yao H, Guin S, Padhye SS, Zhou YQ, Wang MH. Monoclonal antibody (mAb)-induced down-regulation of RON receptor tyrosine kinase diminishes

tumorigenic activities of colon cancer cells. Int J Oncol. 2010; 37(2):473-482. (Clone-specific: Bioassay, Flow cytometry, Fluorescence microscopy, Functional

assay, Immunofluorescence)

Ronsin C, Muscatelli F, Mattei MG, Breathnach R. A novel putative receptor protein tyrosine kinase of the met family. Oncogene. 1993; 8(5):1195-11202. (Biology)

Yao HP, Luo YL, Feng L, et al. Agonistic monoclonal antibodies potentiate tumorigenic and invasive activities of splicing variant of the RON receptor tyrosine

kinase. Cancer Immunol Immunother. 206; 5(9):179-1186. (Immunogen: Bioassay, ELISA, Flow cytometry, Functional assay, Immunohistochemistry,

Immunoprecipitation, Induction, Stimulation)

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