GPNMB Recombinant Rabbit Monoclonal Antibody [PSH0-82]
cat.: HA721511
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH0-82 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 64 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human GPNMB aa 1-498 / 572. |
| Positive control: | L929 cell lysate, A375 cell lysate, U-937 cell lysate, rat heart tissue lysate, B16F1 cell lysates, human lung tissue, L929. |
| Subcellular location: | Cell membrane, Melanosome membrane, Early endosome membrane. |
| Recommended Dilutions:
WB IHC-P IF-Cell FC |
1:1,000 1:200 1:100 1:500-1:1,000 |
| Uniprot #: | SwissProt: Q14956 Human | Q99P91 Mouse | Q6P7C7 Rat |
| Alternative names: | Glycoprotein (transmembrane) nmb Glycoprotein nmb Glycoprotein nmb like protein Gpnmb GPNMB_HUMAN HGFIN NMB Osteoactivin Transmembrane glycoprotein Transmembrane glycoprotein HGFIN Transmembrane glycoprotein NMB |
Images
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Fig1:
Western blot analysis of GPNMB on different lysates with Rabbit anti-GPNMB antibody (HA721511) at 1/1,000 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) Predicted band size: 64 kDa Observed band size: 80-120kDa 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 (HA721511) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of GPNMB on B16F1 cell lysates with Rabbit anti-GPNMB antibody (HA721511) at 1/1,000 dilution. Lysates/proteins at 20 µg/Lane. Predicted band size: 64 kDa Observed band size: 80-120kDa 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 (HA721511) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature. |
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Fig3:
Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-GPNMB antibody (HA721511) 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 (HA721511) 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. |
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Fig4:
Immunocytochemistry analysis of L929 cells labeling GPNMB with Rabbit anti-GPNMB antibody (HA721511) 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 antibody (HA721511) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig5:
Flow cytometric analysis of L929 cells labeling GPNMB. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721511, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.