MA5-12896

antibody from Invitrogen Antibodies

Targeting: NRG1 GGF, HGL, HRG, NDF, NRG1-IT2

Provider product page for MA5-12896

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [29]
Comments [0]
Validations
Western blot [1]
Immunocytochemistry [1]
Immunohistochemistry [1]
Other assay [6]

Submit

Product number: MA5-12896 - Provider product page
Provider: Invitrogen Antibodies
Product name: NRG1 Monoclonal Antibody (7D5)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-12896 targets Heregulin in immunohistochemistry (paraffin), immunoprecipitation, and Western blot applications and shows reactivity with Human, mouse, and Rat samples.
Antibody clone number: 7D5
Concentration: 0.2 mg/mL

NRG1 protein structure - MA5-12896 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1 CSDS induces depression accompanied by an increase in NRG1 degradation. a Experimental timeline of CSDS, SI, behavioural studies and tissue collection. CSDS-induced depression-like behaviours as assessed by b SI test, c the sucrose preference test and d the tail suspension test, and anxiety-like behaviours as assessed by e novelty-suppressed feeding, f the open field test and g the elevated plus maze test, n = 12/12/8. h , i Golgi staining was performed to evaluate changes in dendritic spines in the mPFC between CTR, SS and RES mice, and at least ten neurons from three mice per group were analysed. Scale bars, 10 mum. j - l NRG1 protein level ( n = 8/8/4) was lower in the mPFC of SS mice. There were no differences in NRG1 mRNA levels ( n = 9/8/6) between CTR, SS and RES mice. m - o Experimental timeline of treatment with dexamethasone (Dex), cycloheximide (CHX) and MG132 in primary cortical neurons. Primary cortical neurons were cultured in vitro for a total of 20 days in vitro, including treatment with 0.5 uM Dex twice a day from DIV 10 to DIV 20. Western blot was used to determine levels of NRG1 in primary neurons treated with 10 mug/mL CHX for 4 h, 8 h or 12 h in DIV 20, n = 3 per group. p - r Inhibition of proteasomes by treatment with 10 muM MG132 for 12 h of cultured primary neurons induces NRG1 elevation with Dex treatment for 10 days as measured by western blotting. There were no differences in NRG1 mRNA levels among all groups, n = 3 per group. p -Value signif

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 Overexpression of NRG1 attenuates CSDS-induced depression-like behaviours. a Experimental timeline of mPFC bilateral injection of AAV9-EGFP or AAV9-OE-NRG1 viruses and behavioural studies. b , c Representative western blots and quantification of NRG1 protein in the mPFC of non-stressed (CTR) mice, and mice subjected to CSDS after bilateral injection of AAV9-EGFP or AAV9-OE-NRG1 viruses, n = 3 per group. Overexpression of NRG1 before CSDS-induced depression-like behaviours as assessed by d social interaction, e sucrose preference, f novelty-suppressed feeding, g tail suspension test, h open field test and i the elevated plus maze test, n = 10/7/10/8. p -Value significance is calculated from a one-way ANOVA or two-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001. All data represent mean +- SEM.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 The interaction between Nedd4l and NRG1 promotes the ubiquitination and degradation of NRG1. a Scatter plot of KEGG pathway enrichment from RNA-seq data showed significant changes related to protein degradation in the CTR and SS mice. Highlighted by the red box is the ubiquitin-mediated proteolysis pathway, in which we were most interested. b - d Mice subjected to CSDS showed elevated Nedd4l protein levels in the medial prefrontal cortex (mPFC) of SS mice as measured by western blotting, n = 6 per group. The mRNA level of Nedd4l was enhanced in the mPFC samples of SS mice, and the same result was found with RNA-seq, n = 4/5/3. e - g The mPFC tissue homogenates were immunoprecipitated (IP), using anti-NRG1 and anti-Nedd4l antibodies, n = 4 per group; NRG1 ubiquitination was analysed by western blotting and quantification in CTR, SS and RES mice, n = 3 per group. h - k Representative western blots and quantification of NRG1, Nedd4l and NRG1 ubiquitination (UBI) identified in primary cortical neurons cultured in vitro for a total of 20 days, including treatment with 0.5 uM Dex from DIV 10 to DIV 20, n = 4 per group, two-tailed Student's t -test. l - n Representative western blots and quantification of NRG1 and Nedd4l identified in primary cortical neurons cultured for 20 days, and treated with 1 µL/mL LV-Nedd4l-RNAi virus after treatment with Dex, n = 3 per group. p -Value significance is calculated from a one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001. All d

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 Overexpression of Nedd4l deteriorates subthreshold social defeat stress-induced depression-like behaviours. a Experimental timeline of mPFC bilateral injection of AAV9-EGFP or AAV9-OE-Nedd4l viruses and behavioural studies. Overexpression of Nedd4l before subthreshold social defeat stress (SSDS)-induced depression-like behaviours as assessed by b social interaction, c sucrose preference test and d tail suspension test, and anxiety-like behaviours as assessed by e novelty-suppressed feeding test, f open field test and g the elevated plus maze test, n = 20/9/18/22. h - k Representative western blots and quantification of NRG1, Nedd4l and NRG1 ubiquitination (UBI) identified in the mPFC of non-stressed (CTR) mice, and mice subjected to SSDS after bilateral injection of AAV9-EGFP or AAV9-OE-Nedd4l viruses, n = 3-4 per group. l , m Golgi staining was performed to evaluate the alterations of dendritic spines in the mPFC between CTR and SSDS mice after injection of AAV9-EGFP or AAV9-OE-Nedd4l viruses. At least ten neurons from three mice per group were analysed. Scale bars, 10 mum. p -Value significance is calculated from a one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001. All data represent mean +- SEM.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 5 Knock down of Nedd4l rescues CSDS-induced depression. a Experimental timeline of mPFC bilateral injection of AAV9-EGFP or AAV9-Sh-Nedd4l viruses and behavioural studies. Downregulation of Nedd4l expression before CSDS-induced depression-like behaviours as assessed by b social interaction, c sucrose preference and d tail suspension tests, and anxiety-like behaviours as assessed by e novelty-suppressed feeding, f open field and g the elevated plus maze tests, n = 10/8/8/8. h - k Representative western blots and quantification of NRG1, Nedd4l and NRG1 ubiquitination (UBI) identified in the mPFC of non-stressed (CTR) mice, and mice subjected to CSDS after bilateral injection of AAV9-EGFP or AAV9-Sh-Nedd4l viruses, n = 3 per group. l , m Golgi staining was performed to evaluate the changes of dendritic spines in the mPFC between CTR and CSDS mice after injection of AAV9-EGFP or AAV9-Sh-Nedd4l viruses. At least ten neurons from three mice per group were analysed. Scale bars, 10 mum. p -Value significance is calculated from a one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. All data represent mean +- SEM.