WB
Western blot analysis of GPNMB on different lysates with Rabbit anti-GPNMB antibody at 1/1000 dilution. Lane 1: L929 cell lysate (20 µg/Lane), Lane 2: A375 cell lysate (20 µg/Lane), Lane 3: U-937 cell lysate (20 µg/Lane), Lane 4: Rat heart tissue lysate (40 µg/Lane), Exposure time: 1 minute 55 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100000 dilution was used for 1 hour at room temperature.WB
Western blot analysis of GPNMB on B16F1 cell lysates with Rabbit anti-GPNMB antibody at 1/1000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 10 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of L929 cells labeling GPNMB with Rabbit anti-GPNMB antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-GPNMB antibodyat 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.IHC
Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-GPNMB antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.FC
Flow cytometric analysis of L929 cells labeling GPNMB. Cells were fixed and permeabilized. Then stained with the primary antibody (1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | GPNMB Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description |
|
| Immunogen | Recombinant protein within human GPNMB aa 1-498 / 572. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:1000 ICC/IF:1:100 IHC:1:200 FC:1:500-1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | GPNMB |
|---|---|
| Gene Synonyms | NMB HGFIN PLCA3 |
| Gene Full Name | glycoprotein nmb |
| Gene Summary | 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, Jul 2008] |
| Alternative Names | Glycoprotein (transmembrane) nmb antibody Glycoprotein nmb antibody Glycoprotein nmb like protein antibody Gpnmb antibody GPNMB_HUMAN antibody HGFIN antibody NMB antibody Osteoactivin antibody Transmembrane glycoprotein antibody Transmembrane glycoprotein HGFIN antibody Glycoprotein (transmembrane) nmb antibody Glycoprotein nmb antibody Glycoprotein nmb like protein antibody Gpnmb antibody GPNMB_HUMAN antibody HGFIN antibody NMB antibody Osteoactivin antibody Transmembrane glycoprotein antibody Transmembrane glycoprotein HGFIN antibody Transmembrane glycoprotein NMB antibody |
| Molecular Weight(MW) | 64kDa(Observed band size:80-120kDa) |
| Cellular Localization | Cell membrane, Melanosome membrane, Early endosome membrane. |
WB
Western blot analysis of GPNMB on different lysates with Rabbit anti-GPNMB antibody at 1/1000 dilution. Lane 1: L929 cell lysate (20 µg/Lane), Lane 2: A375 cell lysate (20 µg/Lane), Lane 3: U-937 cell lysate (20 µg/Lane), Lane 4: Rat heart tissue lysate (40 µg/Lane), Exposure time: 1 minute 55 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100000 dilution was used for 1 hour at room temperature.
WB
Western blot analysis of GPNMB on B16F1 cell lysates with Rabbit anti-GPNMB antibody at 1/1000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 10 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of L929 cells labeling GPNMB with Rabbit anti-GPNMB antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-GPNMB antibodyat 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
IHC
Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-GPNMB antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
FC
Flow cytometric analysis of L929 cells labeling GPNMB. Cells were fixed and permeabilized. Then stained with the primary antibody (1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:1000 ICC/IF:1:100 IHC:1:200 FC:1:500-1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
