- PA5-29049 - Invitrogen Antibodies NPR1 antibody

Submitted references Atrial Natriuretic Peptide Promotes Neurite Outgrowth and Survival of Cochlear Spiral Ganglion Neurons in vitro Through NPR-A/cGMP/PKG Signaling.

Sun F, Zhou K, Tian KY, Zhang XY, Liu W, Wang J, Zhong CP, Qiu JH, Zha DJ
Frontiers in cell and developmental biology 2021;9:681421

Natriuretic Peptides Regulate Prostate Cells Inflammatory Behavior: Potential Novel Anticancer Agents for Prostate Cancer.

Mezzasoma L, Talesa VN, Costanzi E, Bellezza I
Biomolecules 2021 May 25;11(6)

Atrial Natriuretic Peptide Improves Neurite Outgrowth from Spiral Ganglion Neurons In Vitro through a cGMP-Dependent Manner.

Sun F, Zhou K, Tian KY, Wang J, Qiu JH, Zha DJ
Neural plasticity 2020;2020:8831735

An approach using Caenorhabditis elegans screening novel targets to suppress tumour cell proliferation.

Mao YQ, Han SF, Zhang SL, Zhang ZY, Kong CY, Chen HL, Li ZM, Cai PR, Han B, Wang LS
Cell proliferation 2020 Jun;53(6):e12832

ANP and BNP Exert Anti-Inflammatory Action via NPR-1/cGMP Axis by Interfering with Canonical, Non-Canonical, and Alternative Routes of Inflammasome Activation in Human THP1 Cells.

Mezzasoma L, Talesa VN, Romani R, Bellezza I
International journal of molecular sciences 2020 Dec 22;22(1)

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of NPR-A using 30 µg of A) H1299 and B) Raji lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a NPR-A polyclonal antibody (Product # PA5-29049) at a dilution of 1:3000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of NPR1 was performed by separating 30 µg of HL-60 whole cell and membrane extracts by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a NPR1 Polyclonal Antibody (Product # PA5-29049) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of NPR1 was achieved by transfecting HeLa cells with NPR1 specific siRNAs (Silencer® select Product # s9686, s9685). Western blot analysis (Fig. a) was performed using membrane extracts from the NPR1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with NPR1 Polyclonal Antibody (Product # PA5-29049, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to NPR1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1) and tissue extracts of Rat Adipose (Lane 2). The blot was probed with Anti-NPR1 Polyclonal Antibody (Product # PA5-29049, 1:1000 dilution) and detected by chemiluminescence using Goat anti Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 118 kDa band corresponding to NPR1 was detected in the cell line and tissue tested. An uncharacterized band was also observed at ~20kDa in the samples tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of NPR-A in paraformaldehyde-fixed HeLa cells using a NPR-A polyclonal antibody (Product # PA5-29049) at a 1:200 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NPR1 Polyclonal Antibody detects NPR1 protein at on HBL435 xenograft by immunohistochemical analysis. Sample: Paraffin-embedded HBL435 xenograft. NPR1 Polyclonal Antibody (Product # PA5-29049) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NPR1 Polyclonal Antibody detects NPR-A protein at cytosol on rat hind brain by immunohistochemical analysis. Sample: Paraffin-embedded rat hind brain. NPR1 Polyclonal Antibody (Product # PA5-29049) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 9 FIGURE NPR1, DCAF4L2 and TSSK6 expression is increased in glioma, and elevated NPR1 or TSSK6 expression is a prognostic indicator of poor survival in patients with classical glioma. A, NPR1, TSSK6 and DCAF4L2 mRNA expression in multiple tumour tissues and normal tissues based on RNA-Seq data from the TCGA. Log2-normalized read count (TPM) is shown. TPM, transcripts per kilobase of exon model per million mapped reads. B, mRNA expression of NPR1, DCAF4L2 and TSSK6 was analysed in glioma tissues from the TCGA data sets. C, Kaplan-Meier survival curve analysis indicated that patients with NPR1 and TSSK6 overexpression had a significantly shorter overall survival in the classical subtype of glioma. D, E, NPR1 protein expression levels were elevated in primary glioma patient samples. F, Immunohistochemical staining of NPR1 in glioma tissue

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Immunolocalization of ANP, NPR-A, and NPR-C in SGNs within the SG of P14 rats. Merge and single-channel images of cochlear sections triple labeled with antibodies against neural marker TUJ1 (green), ANP/NPR-A/NPR-C (red), and DAPI (blue). (a) ANP was predominantly immunoreactive in the perikarya of SGNs. NPR-A (b) and NPR-C (c) were predominantly immunoreactive in the plasma membrane and cytoplasm of SGNs and appeared more pronounced in the cellular membrane. Scale bars = 50 mu m.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Immunolocalization of ANP, NPR-A, and NPR-C in dissociated SGNs from P3 rats. Merge and single-channel images of SG cells triple labeled with antibodies against TUJ1 (green), ANP/NPR-A/NPR-C (red), and DAPI (blue). The immunoreactivity of ANP (a), NPR-A (b), or NPR-C (c) was colocalized with beta -III tubulin-positive SGNs, respectively, distributed in the neuronal soma and neurites. Scale bars = 50 mu m.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Cyclic Guanossine monophosphate (cGMP)/Protein kinase G--I (PKG-I) axis drives Atrial natriuretic peptide (ANP)/ B-type natriuretic peptide (BNP) inhibitory effect on Interleukin (IL)-1beta secretion. THP-1 cells were treated with ANP or BNP (10 -8 M) ( A , B ) or 8-Br-cGMP (100 µM) ( C ) for 1 h ( A , C ) or 30 min ( B ). ( A , C ) NPR-1 protein expression by Western blot. Protein loading was assessed by re-probing the blots with an anti-beta-actin antibody. Western blots are representative of n = 3 independent experiments. ( B ) Cells were lysed, and cGMP intracellular levels were measured by ELISA assay (basal cGMP: 64 fmol/10 6 cells). ( D ) THP-1 cells, either pretreated for 10 min with ANP or BNP (10 -8 M) or 8-Br-cGMP (100 µM) or left untreated, were exposed to LPS (10 µg/mL) for 20 min and treated with ATP (5 mM) for 48 h. When used, KT-5823 (1muM) was added 1 h before ANP, BNP, or cGMP. The supernatants were collected and IL-1beta release was measured by ELISA. All histograms indicate mean +- SD of at least n = three independent experiments, each one tested in triplicate. * p < 0.05, ** p < 0.01, *** p < 0.001 versus untreated control cells; deg p < 0.05, degdeg p < 0.01, versus LPS + ATP-treated cells; # p < 0.05 versus KT-5823 + LPS + ATP treated cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 ANP and BNP counteract PC3-derived EVs-induced inflammasome activation in PNT2 cells. Non-cancerous PNT2 cells were grown for 24 h. Cell lysates were immunoblotted for pro- IL-1beta and mature caspase-1-p20 ( A ); NPR-1, ANP, and BNP ( B ). PNT2 cells were pre-incubated with ANP or BNP (1 uM) for 10 min and then treated with PC3-derived EVs (PC3-EVs) (100 ug/mL) for a 24 h. Cell lysate were immunoblotted for NLRP3 ( C , D ), phospho-NLRP3 (Ser295) ( C , H ), caspase-1 ( C , E ), IL-1beta ( C , F , G ), phospho-ERK 1/2 or ERK 1/2 ( I ) and phospho-p38-MAPK or p38-MAPK ( J ). beta-actin or alpha-tubulin were used as a loading control. Representative Western blots images are shown. Histograms represent densitometric quantification. All histograms indicate the mean +- SD of at least n = 3 independent experiments, each one tested in triplicate. ** p < 0.01, *** p < 0.001 vs. untreated PC3 cells. # p < 0.05, ### p < 0.001 vs. PC3-EVs treated PC3 cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIGURE 2 Immunolocalization of ANP, NPR-A and NPR-C in SGNs within cochlear SG from rats at P7. (A-C) Gross view of cryosectioned cochlear tissues triple-labeled with antibodies against neural marker beta-III Tubulin (TUJ1, green), ANP/NPR-A/NPR-C (red) and Hoechst (blue). (A 1 ,B 1 ,C 1 ) High-magnification views of the boxed region of (A-C) . (A 2 ,B 2 ,C 2 ) Higher-magnification images of the boxed region of (A 1 ,B 1 ,C 1 ) reveal that ANP/NPR-A/NPR-C is colocalized with TUJ1-positive SGNs, respectively. (A 2' ,B 2' ,C 2' ) Immunoreactivities of ANP, NPR-A, and NPR-C are present in the SGNs. Scale bars = 100 mum.

PA5-29049

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