MA5-24014

antibody from Invitrogen Antibodies

Targeting: GPNMB HGFIN, NMB

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-24014

Provider

Invitrogen Antibodies

Product name

GPNMB Monoclonal Antibody (303822)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

In direct ELISAs, no cross-reactivity with recombinant mouse Osteoactivin is observed.

Antibody clone number

303822

Concentration

0.5 mg/mL

References

Submitted references

The suppressive effects of miR-508-5p on the odontogenic differentiation of human dental pulp stem cells by targeting glycoprotein non-metastatic melanomal protein B.

Liu F, Wang X, Yang Y, Hu R, Wang W, Wang Y
Stem cell research & therapy 2019 Jan 22;10(1):35

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of GPNMB was performed using 70% confluent log phase SK-MEL-5 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with GPNMB Monoclonal Antibody (303822) (Product # MA5-24014) at 25 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 488 (Product # A32766, 1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cell membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of GPNMB in MG‚63 human osteosarcoma cell line. Samples were incubated in GPNMB monoclonal antibody (Product # MA5-24014) or isotype control antibody followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody. To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer and permeabilized with Flow Cytometry Permeabilization/Wash Buffer.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 2 The expressions of miR-508-5p and GPNMB during odontogenic differentiation of hDPCs. a miR-508-5p expression levels were detected by qRT-PCR during the odontogenic differentiation of hDPCs at indicated time points. b The expression levels of GPNMB were detected by qRT-PCR during the odontogenic differentiation of hDPCs at indicated time points. c The protein levels of GPNMB were detected by Western blotting during the odontogenic differentiation of hDPCs at indicated time points. d Quantitative analyses of Western blot revealed GPNMB levels increased during the odontogenic differentiation of hDPCs at indicated time points. * p < 0.05 vs the group of 0d

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 3 The 3'-UTR of GPNMB is a direct target of miR-508-5p. a Schematic illustration of the complementary sequence between miR-508-5p and the GPNMB 3'-UTR. The mutation sites on the 3'-UTR of GPNMB were underlined. b hDPCs were transfected with miR-508-5p mimics or siRNA, and the expression levels of GPNMB were determined by qRT-PCR on day 6. c The expression levels of GPNMB were detected by Western blotting after transfection of hDPCs on day 6. d The Western blot results of GPNMB protein were quantified. e Luciferase reporter assay data found that co-transfection of 293T cells with miR-508-5p and wild-type GPNMB 3'-UTR led to a decrease in luciferase activity. In contrast, there was no significant difference observed in the GPNMB-Mt group. * p < 0.05