Anti-GM-CSF receptor antibody cln 2B7-17-A, (monoclonal)

Species Reactivity	Key Applications	Host	Format	Antibody Type
H	FC, IP, WB, NEUT	M	Purified	Monoclonal Antibody

Species Reactivity

Key Applications

Host

Format

Antibody Type

FC, IP, WB, NEUT

Purified

Monoclonal Antibody

Description
Catalogue Number	MAB1037-250UG
Brand Family	Chemicon®
Trade Name	Chemicon
Description	Anti-GM-CSF-α Receptor Antibody, clone K12B7.17A
Alternate Names	CD116

Description

Catalogue Number

MAB1037-250UG

Brand Family

Chemicon®

Trade Name

Chemicon

Description

Anti-GM-CSF-α Receptor Antibody, clone K12B7.17A

Alternate Names

CD116

Product Information
Format	Purified
Presentation	MAB1037 is 0.22 micron filtered in phosphate buffered saline (PBS) with endotoxin levels <0.1ng/μg immunoglobulin as measured by the LAL assay. Purified from ascites fluid by Protein G chromatography and shown to be >95% pure as determined by

Product Information

Format

Purified

Presentation

MAB1037 is 0.22 micron filtered in phosphate buffered saline (PBS) with endotoxin levels <0.1ng/μg immunoglobulin as measured by the LAL assay. Purified from ascites fluid by Protein G chromatography and shown to be >95% pure as determined by

Applications
Application	Detect GM-CSF-α Receptor using this Anti-GM-CSF-α Receptor Antibody, clone K12B7.17A validated for use in FC, IP, WB, NEUT.
Key Applications	Flow Cytometry Immunoprecipitation Western Blotting Neutralizing
Application Notes	Applications include neutralizing the activity of GM CSF and inhibiting human GM-CSF biological action, regardless of whether low-affinity (a-chain only) or high-affinity (a-beta chain heterodimer) receptors are present. The antibody can also be used for Western blot analysis, immunoprecipitation and FACS-flow cytometry. PROTOCOLS: FACS Analysis 1. Incubate 106 cells with 10 μg/mL of antibody at 4°C for 30 min, then wash. 2. Incubate with fluorescein isothiocyanate (FITC) labelled F(ab)2 fragment of sheep ant-mouse Ig antibody at 4°C for 30 min, then wash. Immunoprecipitation 1. Lyse surface iodinated cells (107- 108) in 1% Triton X-100. 2. Incubate the pre-cleared lysate with the antibody at 10 μg/mL for 1 hr at 4°C. 3. Incubate the antibody complexes with Protein-A-Sepharose™ for 1 hr at 4°C. 4. Recover the beads by centrifugation, wash, and then elute with SDS sample buffer prior to running SDS-PAGE. Neutralization The anti GM-CSF receptor (neutralizing) antibody is a potent inhibitor of GM-CSF biological action in several different bioassays with a half maximal inhibitory dose of 1 μg/mL (6 nM) including: 1. Human GM-CSF stimulated 7-day and 14-day bone marrow colonies cultured in agar. 2. Tritiated thymidine incorporation by AML 193 cells. 3. Colony formation by human primary bone marrow cells, and colony formation by FDC-PI cells transfected with the human GM-CSF human receptor a-chain. 4. 125I-human-GM-CSF binding to low-affinity or high-affinity receptors. Western Blot 1. Electroblot SDS-PAGE gels to nitrocellulose sheet and block with 5% w/v milk powder. 2. Incubate nitrocellulose sheet with antibody at 10 μg/mL at room temperature for 1-2 hrs. 3. Incubate nitrocellulose sheet with horseradish peroxidase-labelled rabbit anti-mouse Ig for approximately 1 hr at room temperature. 4. Wash and then develop using Enhanced Chemiluminescence for 5-10 min.

Applications

Application

Detect GM-CSF-α Receptor using this Anti-GM-CSF-α Receptor Antibody, clone K12B7.17A validated for use in FC, IP, WB, NEUT.

Key Applications

Flow Cytometry
Immunoprecipitation
Western Blotting
Neutralizing

Application Notes

Applications include neutralizing the activity of GM CSF and inhibiting human GM-CSF biological action, regardless of whether low-affinity (a-chain only) or high-affinity (a-beta chain heterodimer) receptors are present. The antibody can also be used for Western blot analysis, immunoprecipitation and FACS-flow cytometry.

PROTOCOLS:

FACS Analysis

1. Incubate 106 cells with 10 μg/mL of antibody at 4°C for 30 min, then wash.

2. Incubate with fluorescein isothiocyanate (FITC) labelled F(ab)2 fragment of sheep ant-mouse Ig antibody at 4°C for 30 min, then wash.

Immunoprecipitation

1. Lyse surface iodinated cells (107- 108) in 1% Triton X-100.

2. Incubate the pre-cleared lysate with the antibody at 10 μg/mL for 1 hr at 4°C.

3. Incubate the antibody complexes with Protein-A-Sepharose™ for 1 hr at 4°C.

4. Recover the beads by centrifugation, wash, and then elute with SDS sample buffer prior to running SDS-PAGE.

Neutralization

The anti GM-CSF receptor (neutralizing) antibody is a potent inhibitor of GM-CSF biological action in several different bioassays with a half maximal inhibitory dose of 1 μg/mL (6 nM) including:

1. Human GM-CSF stimulated 7-day and 14-day bone marrow colonies cultured in agar.

2. Tritiated thymidine incorporation by AML 193 cells.

3. Colony formation by human primary bone marrow cells, and colony formation by FDC-PI cells transfected with the human GM-CSF human receptor a-chain.

4. 125I-human-GM-CSF binding to low-affinity or high-affinity receptors.

Western Blot

1. Electroblot SDS-PAGE gels to nitrocellulose sheet and block with 5% w/v milk powder.

2. Incubate nitrocellulose sheet with antibody at 10 μg/mL at room temperature for 1-2 hrs.

3. Incubate nitrocellulose sheet with horseradish peroxidase-labelled rabbit anti-mouse Ig for approximately 1 hr at room temperature.

4. Wash and then develop using Enhanced Chemiluminescence for 5-10 min.

Biological Information
Clone	K12B7.17A
Host	Mouse
Specificity	MAB1037 neutralizes the human native granulocyte-macrophage colony stimulating factor (GM-CSF) receptor and recombinant GM-CSF receptor. Binds to the alpha chain of human GM-CSF receptor inhibiting both high-affinity (alpha-beta chain heterodimer) and low-affinity (alpha chain only) binding and subsequent action of GM-CSF(Nicola, 1993). This antibody has a relatively high-affinity for the GM-CSF receptor alpha chain (KD-3nM).
Isotype	IgG2a
Species Reactivity	Human
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NM_172248.1 NM_006140.3 NM_172245.1 NM_172246.1 NM_172247.1 NM_172249.1
Entrez Gene Summary	The protein encoded by this gene is the alpha subunit of the heterodimeric receptor for colony stimulating factor 2, a cytokine which controls the production, differentiation, and function of granulocytes and macrophages. The encoded protein is a member of the cytokine family of receptors. This gene is found in the pseudoautosomal region (PAR) of the X and Y chromosomes. Multiple transcript variants encoding different isoforms have been found for this gene, with some of the isoforms being membrane-bound and others being soluble.
Gene Symbol	CSF2RA CSF2R CSF2RAY GMR CSF2RX MGC4838 GM-CSF-R-alpha CSF2RAX CD116 MGC3848 CSF2RY CDw116 GMCSFR
UniProt Number	P15509
UniProt Summary	FUNCTION: SwissProt: P15509 # Low affinity receptor for granulocyte-macrophage colony- stimulating factor. Transduces a signal that results in the proliferation, differentiation, and functional activation of hematopoietic cells. SIZE: 400 amino acids; 46207 Da SUBUNIT: Heterodimer of an alpha and a beta chain. The beta chain is common to the IL3, IL5 and GM-CSF receptors. SUBCELLULAR LOCATION: Cell membrane; Single-pass type I membrane protein. & Isoform 3: Secreted (Probable). & Isoform 4: Secreted (Probable). & Isoform 6: Secreted (Probable). DOMAIN: SwissProt: P15509 The WSXWS motif appears to be necessary for proper protein folding and thereby efficient intracellular transport and cell- surface receptor binding. & The box 1 motif is required for JAK interaction and/or activation. SIMILARITY: Belongs to the type I cytokine receptor family. Type 5 subfamily. MISCELLANEOUS: The gene encoding for this protein is located in the pseudoautosomal region 1 (PAR1) of X and Y chromosomes.

Biological Information

Clone

K12B7.17A

Host

Mouse

Specificity

MAB1037 neutralizes the human native granulocyte-macrophage colony stimulating factor (GM-CSF) receptor and recombinant GM-CSF receptor. Binds to the alpha chain of human GM-CSF receptor inhibiting both high-affinity (alpha-beta chain heterodimer) and low-affinity (alpha chain only) binding and subsequent action of GM-CSF(Nicola, 1993). This antibody has a relatively high-affinity for the GM-CSF receptor alpha chain (KD-3nM).

Isotype

IgG2a

Species Reactivity

Human

Antibody Type

Monoclonal Antibody

Entrez Gene Number

Entrez Gene Summary

The protein encoded by this gene is the alpha subunit of the heterodimeric receptor for colony stimulating factor 2, a cytokine which controls the production, differentiation, and function of granulocytes and macrophages. The encoded protein is a member of the cytokine family of receptors. This gene is found in the pseudoautosomal region (PAR) of the X and Y chromosomes. Multiple transcript variants encoding different isoforms have been found for this gene, with some of the isoforms being membrane-bound and others being soluble.

Gene Symbol

CSF2RA
CSF2R
CSF2RAY
GMR
CSF2RX
MGC4838
GM-CSF-R-alpha
CSF2RAX
CD116
MGC3848
CSF2RY
CDw116
GMCSFR

UniProt Number

P15509

UniProt Summary

FUNCTION: SwissProt: P15509 # Low affinity receptor for granulocyte-macrophage colony- stimulating factor. Transduces a signal that results in the proliferation, differentiation, and functional activation of hematopoietic cells.
SIZE: 400 amino acids; 46207 Da
SUBUNIT: Heterodimer of an alpha and a beta chain. The beta chain is common to the IL3, IL5 and GM-CSF receptors.
SUBCELLULAR LOCATION: Cell membrane; Single-pass type I membrane protein. & Isoform 3: Secreted (Probable). & Isoform 4: Secreted (Probable). & Isoform 6: Secreted (Probable).
DOMAIN: SwissProt: P15509 The WSXWS motif appears to be necessary for proper protein folding and thereby efficient intracellular transport and cell- surface receptor binding. & The box 1 motif is required for JAK interaction and/or activation.
SIMILARITY: Belongs to the type I cytokine receptor family. Type 5 subfamily.
MISCELLANEOUS: The gene encoding for this protein is located in the pseudoautosomal region 1 (PAR1) of X and Y chromosomes.

Product Usage Statements
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Anti-GM-CSF receptor (neutralizing) is shipped liquid at ambient temperature. Store at 2-8°C for up to 6 months.

Storage and Shipping Information

Storage Conditions

Anti-GM-CSF receptor (neutralizing) is shipped liquid at ambient temperature. Store at 2-8°C for up to 6 months.

Packaging Information
Material Size	250 µg

Packaging Information

Material Size

250 µg

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

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Anti-GM-CSF receptor antibody cln 2B7-17-A, (monoclonal)

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