Osteoactivin antibody (AA 81-180)

Catalog No. ABIN749093

Product Details anti-Osteoactivin Antibody

Target: Osteoactivin (GPNMB)

Binding Specificity: AA 81-180

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This Osteoactivin antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunofluorescence (Cultured Cells) (IF (cc)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunohistochemistry (Frozen Sections) (IHC (fro))

Cross-Reactivity: Human, Mouse, Rat

Predicted Reactivity: Cow,Pig,Horse,Guinea Pig

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human GPNMB

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Restrictions: For Research Use only

Independent Validation (IHC (fp))

Validation #100002 (Immunohistochemistry (Formalin-fixed Paraffin-embedded Sections))

by: Memorial Sloan-Kettering Cancer Center, Parada lab

No.: #100002

Date: 10/15/2015

Antigen: Osteoactivin

Method validated: Immunohistochemistry (Formalin-fixed Paraffin-embedded Sections)

Positive Control: Tumor cell transplanted mouse brain was dissected

Notes: The staining pattern was as expected for Osteoactivin as membrane protein.

Validation Images

IF validation of osteoactivin antibody ABIN749093 on mouse brain tumor sections. A. ABIN749093 diluted 1:100 (red), DAPI counterstaining (blue). B. Dilution factor 1:50. C. Dilution factor 1:100. D. Dilution factor 1:200.

Full Methods

Primary Antibody: ABIN749093

Secondary Antibody: Donkey-anti-Rabbit-Cy3

Full Protocol:

• Tumor cell transplanted mouse brain was dissected, fixed in 4% PFA for overnight, saturated in 30% sucrose, mounted in Cryo, and sectioned into 12µm sections.
• Immunostaining for Cryo Sections:
• Dry slides at 50-55°C for 15-30 min (or RT for 30min)
• Wash three times with PBS 5min (shake gently)
• Block with 10% Normal Donkey Serum (NDS) at RM for 1hour
• Incubate with primary antibody at 4°C overnight (Prepared in 2% NDS)
• Rabbit-anti-Osteoactivin antibody (ABIN749093) diluted 1:50-1:400
• Wash three times with PBS 5min (shake gently)
• Incubate with secondary antibody together with DAPI at RT for 1h (Prepared in 2% NDS), cover the chamber with foil
• Donkey-anti-Rabbit-Cy3
• DAPI 1:100
• Wash three times with PBS 5min (shake gently)
• Add three drops of water-based Immu-mount
• Cover with 24x60 cover glass
• Imaging
• Images were taken using a Zeiss Axio2-Imaging system, 20X-0.5NA-Dry Dye-1004-072 objective, DAPI and Cy3 channel.

Experimental Notes: ABIN749093 worked within the diution range 1:50 - 1:200; the antibody did not work well at a 1:400 dilution.

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-Osteoactivin antibody (ABIN749093)

Neal, Boyle, Budge, Safadi, Richardson: "The glycoprotein GPNMB attenuates astrocyte inflammatory responses through the CD44 receptor." in: Journal of neuroinflammation, Vol. 15, Issue 1, pp. 73, (2019) (PubMed).

Oyewumi, Manickavasagam, Novak, Wehrung, Paulic, Moussa, Sondag, Safadi: "Osteoactivin (GPNMB) ectodomain protein promotes growth and invasive behavior of human lung cancer cells." in: Oncotarget, Vol. 7, Issue 12, pp. 13932-44, (2017) (PubMed).

Arosarena, Dela Cadena, Denny, Bryant, Barr, Thorpe, Safadi: "Osteoactivin Promotes Migration of Oral Squamous Cell Carcinomas." in: Journal of cellular physiology, Vol. 231, Issue 8, pp. 1761-70, (2016) (PubMed).

Frara, Abdelmagid, Tytell, Amin, Popoff, Safadi, Barbe: "Growth and repair factors, osteoactivin, matrix metalloproteinase and heat shock protein 72, increase with resolution of inflammation in musculotendinous tissues in a rat model of repetitive grasping." in: BMC musculoskeletal disorders, Vol. 17, pp. 34, (2016) (PubMed).

Ruan, Li, Nie, Zhou, Zou: "Differential expression of glycoprotein non-metastatic melanoma protein B (GPNMB) involved in trichostatin A-induced apoptosis in gastric cancer." in: International journal of clinical and experimental medicine, Vol. 7, Issue 12, pp. 4857-66, (2015) (PubMed).

Abdelmagid, Sondag, Moussa, Belcher, Yu, Stinnett, Novak, Mbimba, Khol, Hankenson, Malcuit, Safadi: "Mutation in Osteoactivin Promotes Receptor Activator of NF?B Ligand (RANKL)-mediated Osteoclast Differentiation and Survival but Inhibits Osteoclast Function." in: The Journal of biological chemistry, Vol. 290, Issue 33, pp. 20128-46, (2015) (PubMed).

Target Details for Osteoactivin

Target: Osteoactivin (GPNMB)

Alternative Name: GPNMB (GPNMB Products)

Synonyms: HGFIN antibody, NMB antibody, DC-HIL antibody, Dchil antibody, ipd antibody, glycoprotein nmb antibody, glycoprotein (transmembrane) nmb antibody, glycoprotein nmb L homeolog antibody, Gpnmb antibody, GPNMB antibody, gpnmb.L antibody

Background:

Synonyms: NMB, HGFIN, Transmembrane glycoprotein NMB, Transmembrane glycoprotein HGFIN, GPNMB, UNQ1725/PRO9925

Background: The protein encoded by this gene is a type I transmembrane glycoprotein which shows homology to the pMEL17 precursor, a melanocyte-specific protein. GPNMB shows expression in the lowly metastatic human melanoma cell lines and xenografts but does not show expression in the highly metastatic cell lines. GPNMB may be involved in growth delay and reduction of metastatic potential. Two transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq].

Gene ID: 10457

UniProt: Q14956