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AB317325

Anti-GPNMB antibody [EPR29063-10]

  • BOND RX™ Validated
  • 20ul selling size
  • Recombinant
  • RabMAb
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Rabbit Recombinant Monoclonal GPNMB antibody. Suitable for WB, ICC/IF, IHC-P and reacts with Mouse, Rat samples.

View Alternative Names

Dchil, Hgfin, Nmb, Gpnmb, Transmembrane glycoprotein NMB, DC-HIL, Dendritic cell-associated transmembrane protein, Osteoactivin

9 Images
Immunocytochemistry/ Immunofluorescence - Anti-GPNMB antibody [EPR29063-10] (AB317325)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GPNMB antibody [EPR29063-10] (AB317325)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized B16-F0 (mouse melanoma epithelial-like cell) cells labelling GPNMB with ab317325 at 1/500 (1.036 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in B16-F0 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control : EL4 (PMID : 19320736).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR29063-10] (AB317325)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR29063-10] (AB317325)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling GPNMB with ab317325 at 1/5000 (0.104 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat spleen.
The section was incubated with ab317325 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR29063-10] (AB317325)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR29063-10] (AB317325)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling GPNMB with ab317325 at 1/5000 (0.104 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen.
The section was incubated with ab317325 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR29063-10] (AB317325)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR29063-10] (AB317325)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling GPNMB with ab317325 at 1/5000 (0.104 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : scattered staining on immune cells of mouse testis.
The section was incubated with ab317325 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR29063-10] (AB317325)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR29063-10] (AB317325)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling GPNMB with ab317325 at 1/5000 (0.104 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : scattered staining on immune cells of rat testis.
The section was incubated with ab317325 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR29063-10] (AB317325)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR29063-10] (AB317325)

Immunohistochemical analysis of paraffin-embedded A) B16-F0 (mouse melanoma epithelial-like cell) cell pellet; B) EL4 (mouse lymphoma T lymphocyte) cell pellet tissue labeling GPNMB with ab317325 at 1/5000 (0.104 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on A) B16-F0 cell pellet; no staining on B) EL4 cell pellet.
The section was incubated with ab317325 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR29063-10] (AB317325)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR29063-10] (AB317325)

Immunohistochemical analysis of paraffin-embedded Mouse lung cancer tissue labeling GPNMB with ab317325 at 1/5000 (0.104 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung cancer (PMID : 29857071).
The section was incubated with ab317325 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Western blot - Anti-GPNMB antibody [EPR29063-10] (AB317325)
  • WB

Supplier Data

Western blot - Anti-GPNMB antibody [EPR29063-10] (AB317325)

Negative control : EL4(PMID : 19320736), 4T1(PMID : 20711474)

The identity of the lower MW band at approximately 15 kDa (in lane1 and 2) is unknown.

The molecular mass observed is consistent with what has been described in the literature (PMID : 19320736).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-GPNMB antibody [EPR29063-10] (ab317325) at 1/1000 dilution

Lane 1:

B16-F10 (Mouse skin melanoma) whole cell lysate at 20 µg

Lane 2:

B16-F0 (mouse melanoma epithelial cell-like) whole cell lysate at 20 µg

Lane 3:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Lane 4:

4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 95 kDa,120 kDa,36 kDa

false

Exposure time: 37s

Western blot - Anti-GPNMB antibody [EPR29063-10] (AB317325)
  • WB

Supplier Data

Western blot - Anti-GPNMB antibody [EPR29063-10] (AB317325)

Low expression : testis

The molecular mass observed is consistent with what has been described in the literature (PMID : 19320736).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-GPNMB antibody [EPR29063-10] (ab317325) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Lane 3:

Rat testis tissue lysate at 20 µg

Lane 4:

Rat spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 65 kDa,120 kDa,36 kDa

false

Exposure time: 180s

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR29063-10

Isotype

IgG

Carrier free

No

Reacts with

Mouse, Rat

Applications

ICC/IF, IHC-P, WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Mouse": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/500", "ICCIF-species-notes": "<p></p>", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/5000", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p></p>", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>" }, "Rat": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/5000", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" } } }
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Product details

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

GPNMB also known as osteoactivin is a type I transmembrane glycoprotein with a molecular mass of approximately 115 kDa. This protein is highly expressed in various tissues including the bone skin and brain. It also shows notable presence in certain immune cells such as dendritic cells and macrophages. GPNMB localizes mainly on the cell membrane and within lysosomal compartments indicating its involvement in cellular and signaling processes.
Biological function summary

GPNMB plays versatile roles in promoting cell growth differentiation and repair. It is involved in processes like tissue regeneration possibly due to its role in cell adhesion and migration. The protein also partakes in osteoclast differentiation and bone resorption therefore linking to skeletal homeostasis. GPNMB forms part of a broader protein complex at the cellular junctions interacting with integrins to mediate its functions.

Pathways

GPNMB features prominently in osteoclastogenesis and melanoma metastasis pathways. It cooperates with RANKL an important protein in bone metabolism by influencing osteoclast differentiation and activation. In the context of melanoma GPNMB's action in pathways may enhance tumor proliferation and invasion possibly interacting with signaling molecules like AKT or ERK although the specifics remain actively researched.

GPNMB has significant links to both osteoarthritis and certain types of cancer particularly breast cancer and melanoma. In osteoarthritis GPNMB expression often increases reflecting involvement in cartilage degradation processes. Its overexpression correlates with tumor progression in melanoma potentially driven by its interaction with integrin β1 which assists in cellular adhesion and migration within the tumor microenvironment.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Could be a melanogenic enzyme.
See full target information Gpnmb
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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