Invitrogen

Phospho-DRP1 (Ser616) Polyclonal Antibody

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This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

6 Published Figures

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Phospho-DRP1 (Ser616) Antibody in Western Blot (WB) — Western blot was performed using Anti-Phospho-DRP1 (Ser616) Polyclonal Antibody (Product # PA5-64821) and 58, 74 kDa bands corresponding to Phospho-DRP1 (Ser616) were observed (Fig. A). Whole cell extracts (30 µg lysate) of IMR-32 (Lane 1), SH-SY5Y (Lane 2) and Daudi (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). For Fig. B, the blot was treated with Lambda Protein Phosphatase after the protein transfer and for Fig. A, the blot was left untreated. In Fig.a, band of interest is not present in Daudi, a low expressing model for DRP1. Phospho-specificity is demonstrated by absence of band in Fig.b.

Phospho-DRP1 (Ser616) Antibody in Western Blot (WB) — Western blot analysis of DRP1 in UV treated 293 whole cell lysates using a Phospho-DRP1 (Ser616) Polyclonal Antibody (Product # PA5-64821).

Phospho-DRP1 (Ser616) Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across the cell lines tested owing to their inherent genetic constitution. Relative expression of Phospho-DRP1 (Ser616) was observed in IMR32, SH-SY5Y in comparison to Daudi, a model that expresses low levels of DRP1 using Anti-Phospho-DRP1 (Ser616) Polyclonal Antibody (Product # PA5-64821) in Western Blot. {RE}

Phospho-DRP1 (Ser616) Antibody — TGF-beta1 induces mitochondrial fusion in vascular progenitor cells (VPCs). A, Representative images of mitochondrial morphology determined by MitoTracker staining in control-VPCs and Marfan syndrome (MFS)-VPCs. B, The mitochondrial length in control-VPCs and MFS-VPCs was analysed. C, Western blotting and quantitative analysis of the level of p-Drp1 and Mfn2 protein in control-VPCs and MFS-VPCs. D, Western blotting and quantitative analysis of the level of p-Drp1 and Mfn2 protein in control-VPCs that received TGF-beta1 or TGF-beta1 + Mfn2-siRNA treatment. E, Representative images of SA-beta-gal staining in control-VPCs that received TGF-beta1 or TGF-beta1 + Mfn2-siRNA treatment. F, The SA-beta-gal positive cells in control-VPCs that received TGF-beta1 or TGF-beta1 + Mfn2-siRNA treatment were calculated and are presented as a percentage of total cells. G, Representative images of Mito-Sox staining in control-VPCs that received TGF-beta1 or TGF-beta1 + Mfn2-siRNA treatment. H, Quantitative analysis of ROS generation in control-VPCs that received TGF-beta1 or TGF-beta1 + Mfn2-siRNA treatment. Data are expressed as mean +- SEM (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bar = 100 mum {TM}

Phospho-DRP1 (Ser616) Antibody in Western Blot (WB) — FIGURE 5 Effect of drug treatment on the expression of proteins associated with mitochondrial function and AMPK signaling in CIA-induced rats. Sprague-Dawley rats were induced by CIA for 14 days and subjected to daily drug administration at various doses for 20 consecutive days. (A) Western blot detection and quantification of the expression of mitochondrial function-related proteins (Drp1, Drp1 phosphorylation at Ser616 and Ser637, MTFP1, and MFN2) in synovial tissues. (B) Western blot detection and quantification of the expression of AMPK in its non-phosphorylated and phosphorylated form in synovial tissues. All protein expression was normalized to that of GAPDH as a housekeeping control. All data are expressed as the mean +- standard deviation ( n = 3). * p < 0.05. Low, medium, and high doses of COR and PAE were administered, and dose response was analyzed. CIA: collagen-induced arthritis; COR: Cornus officinalis; PAE: Paeonia lactiflora; PF: paeoniflorin; UA: ursolic acid; DEX: dexamethasone; Drp1: dynamin-related protein 1; MTFN1: mitochondrial fission process protein 1; MFN2: mitofusin 2; AMPK: AMP-activated protein kinase; GAPDH: glyceraldehyde 3-phosphate dehydrogenase.

Phospho-DRP1 (Ser616) Antibody in Western Blot (WB) — FIGURE 2 Acute kidney injury (AKI) induces damage to cardiomyocyte mitochondria. (A) ATP production was measured through Enhanced ATP Assay Kit. (B,C) Cardiomyocytes were isolated from mice after AKI. Then, mitochondrial membrane potential was determined through JC-1 probe in primary cardiomyocytes. The ratio of red-to-green fluorescence intensity was used to evaluate mitochondrial membrane potential. (D,E) In cardiomyocytes, DCFH-DA was used to observe the levels of intracellular reactive oxygen species. (F,G) Immunofluorescence of the mitochondria. The number of fragmented mitochondria in cardiomyocytes was recorded. (H,I) Western blots were used to analyze the expression of phosphorylated Drp1 (p-Drp1). The relative expression of p-Drp1 was evaluated in cardiomyocytes. * p < 0.05 vs. sham group.

Phospho-DRP1 (Ser616) Antibody in Western Blot (WB) — FIGURE 5 RH impairs mitochondrial function and promotes excessive Drp1-mediated fission via repressing TRPC6 in diabetic mice. (A) Summarized data for the oxygen consumption capacity of hippocampal mitochondria measured by high-resolution respirometry (left) and ATP content (right) of hippocampus from diabetic WT or TRPC6 -/- mice with or without RH (n = 6 tissues from mice). CI Leak, leak respiration of CI; CI OXPHOS, CI + CII OXPHOS, CI, and CI plus CII oxidative phosphorylation capacity; CI ETS, CI + CII ETS, CI, and CI plus CII electron transfer system capacity; CI ETS, CI + CII ETS, CI, and CI plus CII electron transfer system capacity. (B) Summarized data for the oxygen consumption capacity of mitochondria (left) and ATP content (right) of hippocampal from hyperforin or vehicle-treated diabetic WT or TRPC6 -/- mice with RH (n = 6 tissues from 6 mice). (C and D) TEM images of mitochondria (indicated by yellow arrows) in the CA1 region of hippocampus. Quantitative data of mitochondrial length and ratio of length to width are shown below (n = 80-120 mitochondria from three mice). Scale bar, 1 um. (E, F) Western blots of MFN1, MFN2, Drp1, p-Drp1 (S616), and p-Drp1 (S637) in hippocampus homogenates of mice. Quantitative data are shown on the right (n = 4 tissues from four mice). * P < .05, ** P < .01, *** P < .01 (two-way or one-way ANOVA or Kruskal-Wallis test)

Phospho-DRP1 (Ser616) Antibody in Western Blot (WB) — Fig. 6 Phosphorylation of RIPK3 in serine 204 regulates mitochondrial fission via DRP1. ( a ) IF staining of phosphorylated RIPK3 (pRIPK3) in murine aortic sections of animals treated as indicated (scale bar = 20 um). ( b ) Hematoxylin/Eosin (H&E) and IF staining of pRIPK3 (red) and CD68 (green) in healthy or aneurysmal human tissue (scale bar H&E = 500 um, IF = 20 um). Merge is shown in yellow and nucleus is stained with DAPI in blue. Dotted lines define the magnified region. L lumen. ( c ) Immunoblot of pRIPK3 and quantification of healthy or aneurysmal human tissue. n = 3 per group. ** P = 0.0037. ( d ) IF staining of pRIPK3 in BMDM stimulated with HMGB1 (10 ng mL -1 ). Nucleus is stained with DAPI in blue (scale bar = 10 um). ( e ) Schematic representation of RIPK3 phosphorylation sites. Targeted mutations designed in synthetic modified RNA (modRNA) are illustrated. WT, wild type; S, serine; A, Alanine; N-T, N-terminal; C-T, C-terminal; S204>A, modification of serine 204; S232>A modification of serine 232; DM, double serine mutations. ( f ) Representative histograms of flow cytometry analysis of Mitotracker and quantification of MFI in BMDM transfected with Ripk3 modRNA as indicated. n = 4 (DM) and 5 (Control, WT, 204>A and 232>A). * P < 0.05, ** P < 0.01. Error bars represent s.e.m. ( g ) Immunoblot analysis and quantification of pDRP1 in BMDM transfected with Ripk3 modRNA as indicated. n = 3 . * P < 0.05, ** P < 0.01. Data is presented as mean, error bars represent s.e.

Phospho-DRP1 (Ser616) Antibody in Western Blot (WB) — Fig. 5 HMGB1-induced RIPK3 promotes mitochondrial fission in macrophages. Oxygen consumption rate (OCR) and respiration quantifications of WT and Ripk3 -/- BMDM treated with HMGB1 ( a , b ). n = 3 (WT + HMGB1, Ripk3 -/- and Ripk3 -/- +HMGB1) and 7 (WT). * P < 0.05, *** P < 0.001. OCR profiles and quantification of WT and Ripk3 -/- BMDM treated with HMGB1 in the presence of GSK'872 (10 µM, 1 h) ( c , d ). n = 5 per group. ** P < 0.01, *** P < 0.001. Representative histograms of Mitotracker and quantification of the mean fluorescence intensity (MFI) of WT or Ripk3 -/- BMDM treated with HMGB1 ( e ). n = 3 ( Ripk3 -/- and Ripk3 -/- +HMGB1), 5 (WT + HMGB1) and 6 (WT). * P < 0.05, *** P < 0.001. Representative histograms of Mitotracker and quantification of MFI of WT BMDM treated with lung extracts of PBS-treated mice (CTL) or ALI lung extracts (ALI) pre-incubated with anti-HMGB1 (alpha-HMGB1) or with control IgG ( f ). n = 3 per group. * P < 0.05. ( g ) Representative electron microscopy images of mitochondria in WT or Ripk3 -/- BMDM treated with HMGB1 or PBS (CTL). Scale bar = 500 nm. Quantification of mitochondria number per cell ( h ) in WT or Ripk3 -/- as indicated. n = 11 (WT), 10 (WT + HMGB1), 12 ( Ripk3 -/- ) and (17 Ripk3 -/- +HMGB1). ** P < 0.01. ( i ) IF staining of Mitotracker and quantification of mean mitochondrial area in WT or Ripk3 -/- BMDM treated with HMGB1 (10 ng mL -1 ). Scale bar = 10 um. n = 4 per group. * P = 0.0358. ( j ) Immunoblot of pDRP1 and quant

Phospho-DRP1 (Ser616) Antibody in Western Blot (WB) — Figure 4 Mitochondrial function was decreased in AAA-ASCs. (a) Representative images of mitochondrial morphology determined by MitoTracker staining in H-ASCs and AAA-ASCs. Scale bar = 25 mu m. (b) Western blotting and quantitative analysis of the expression level of p-Drp1 ser616/Drp1 and Mfn2 in H-ASCs and AAA-ASCs. (c) Representative images of ROS determined by MitoSOX staining and quantitative analysis of ROS generation in H-ASCs and AAA-ASCs. Scale bar = 100 mu m. (d) Representative images of MMP determined by JC-1 staining and quantitative analysis of MMP in H-ASCs and AAA-ASCs. (e) The intracellular ATP level in H-ASCs and AAA-ASCs. Data are expressed as mean +- SEM. n = 3. *** p < 0.001.

Product Details

PA5-64821

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000	View 10 publications 10 publications
Immunohistochemistry (IHC)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Mouse, Rat
Published species	Human, Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	A synthesized peptide derived from human DRP1 (Phospho-Ser616) View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Affinity chromatography
Storage buffer	PBS, pH 7.4, with 50% glycerol
Contains	0.02% sodium azide
Storage conditions	-20°C
RRID	AB_2663284

Product Specific Information

Phospho-DRP1 (Ser616) Polyclonal Antibody detects endogenous levels of DRP1 only when phosphorylated at Ser616.

Target Information

A human dynamin-related protein, DRP1 contributes to mitochondrial division in mammalian cells. It plays this important role in mitochondrial fission at steady state and during apoptosis. DRP1 is required for proper cellular distribution of mitochondria, and in mutant neurons, mitochondria are largely absent from synapses, thus providing a genetic tool to assess the role of mitochondria at synapses.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

DRP1 Promotes BRAF^V600E-Driven Tumor Progression and Metabolic Reprogramming in Colorectal Cancer.

Frontiers in oncology

Padder RA,Bhat ZI,Ahmad Z,Singh N,Husain M

PA5-64821 was used in Western Blot to investigate how BRAFV600E driven CRC (colorectal cancer) progression is linked to mitochondrial fission/fusion dynamics and whether this window could be exploited to target CRC progression.

Thu Apr 21 00:00:00 EDT 2022

Combined Cornus Officinalis and Paeonia Lactiflora Pall Therapy Alleviates Rheumatoid Arthritis by Regulating Synovial Apoptosis via AMPK-Mediated Mitochondrial Fission.

Frontiers in pharmacology

Huang L,Hu S,Shao M,Wu X,Zhang J,Cao G

Published figure using Phospho-DRP1 (Ser616) polyclonal antibody (Product # PA5-64821) in Western Blot

Wed Apr 28 00:00:00 EDT 2021

Grb2 Induces Cardiorenal Syndrome Type 3: Roles of IL-6, Cardiomyocyte Bioenergetics, and Akt/mTOR Pathway.

Frontiers in cell and developmental biology

Wang J,Sun X,Wang X,Cui S,Liu R,Liu J,Fu B,Gong M,Wang C,Shi Y,Chen Q,Cai G,Chen X

Published figure using Phospho-DRP1 (Ser616) polyclonal antibody (Product # PA5-64821) in Western Blot

Fri Apr 09 00:00:00 EDT 2021

Low-glucose-sensitive TRPC6 dysfunction drives hypoglycemia-induced cognitive impairment in diabetes.

Clinical and translational medicine

He C,Gao P,Cui Y,Li Q,Li Y,Lu Z,Ma H,Zhao Y,Li L,Sun F,Chen X,Jia H,Liu D,Yang G,Zheng H,Zhu Z

Published figure using Phospho-DRP1 (Ser616) polyclonal antibody (Product # PA5-64821) in Western Blot

Thu Oct 01 00:00:00 EDT 2020

Low-glucose-sensitive TRPC6 dysfunction drives hypoglycemia-induced cognitive impairment in diabetes.

Clinical and translational medicine

He C,Gao P,Cui Y,Li Q,Li Y,Lu Z,Ma H,Zhao Y,Li L,Sun F,Chen X,Jia H,Liu D,Yang G,Zheng H,Zhu Z

Published figure using Phospho-DRP1 (Ser616) polyclonal antibody (Product # PA5-64821) in Western Blot

Thu Oct 01 00:00:00 EDT 2020

Lung-derived HMGB1 is detrimental for vascular remodeling of metabolically imbalanced arterial macrophages.

Nature communications

Boytard L,Hadi T,Silvestro M,Qu H,Kumpfbeck A,Sleiman R,Fils KH,Alebrahim D,Boccalatte F,Kugler M,Corsica A,Gelb BE,Jacobowitz G,Miller G,Bellini C,Oakes J,Silvestre JS,Zangi L,Ramkhelawon B

Published figure using Phospho-DRP1 (Ser616) polyclonal antibody (Product # PA5-64821) in Western Blot

Thu Aug 27 00:00:00 EDT 2020

Lung-derived HMGB1 is detrimental for vascular remodeling of metabolically imbalanced arterial macrophages.

Nature communications

Boytard L,Hadi T,Silvestro M,Qu H,Kumpfbeck A,Sleiman R,Fils KH,Alebrahim D,Boccalatte F,Kugler M,Corsica A,Gelb BE,Jacobowitz G,Miller G,Bellini C,Oakes J,Silvestre JS,Zangi L,Ramkhelawon B

Published figure using Phospho-DRP1 (Ser616) polyclonal antibody (Product # PA5-64821) in Western Blot

Thu Aug 27 00:00:00 EDT 2020

Lung-derived HMGB1 is detrimental for vascular remodeling of metabolically imbalanced arterial macrophages.

Nature communications

Boytard L,Hadi T,Silvestro M,Qu H,Kumpfbeck A,Sleiman R,Fils KH,Alebrahim D,Boccalatte F,Kugler M,Corsica A,Gelb BE,Jacobowitz G,Miller G,Bellini C,Oakes J,Silvestre JS,Zangi L,Ramkhelawon B

Published figure using Phospho-DRP1 (Ser616) polyclonal antibody (Product # PA5-64821) in Western Blot

Thu Aug 27 00:00:00 EDT 2020

Adipose-Derived Mesenchymal Stem Cells Isolated from Patients with Abdominal Aortic Aneurysm Exhibit Senescence Phenomena.

Oxidative medicine and cellular longevity

Huang X,Zhang H,Liang X,Hong Y,Mao M,Han Q,He H,Tao W,Jiang G,Zhang Y,Li X

Published figure using Phospho-DRP1 (Ser616) polyclonal antibody (Product # PA5-64821) in Western Blot

Fri May 01 00:00:00 EDT 2020

FGF21 Mediates Mesenchymal Stem Cell Senescence via Regulation of Mitochondrial Dynamics.

Oxidative medicine and cellular longevity

Li X,Hong Y,He H,Jiang G,You W,Liang X,Fu Q,Han S,Lian Q,Zhang Y

PA5-64821 was used in Western Blotting to study the mechanisms of fibroblast growth factor 21 in the regulation of mesenchymal stem cell senescence.

Thu Jan 16 00:00:00 EST 2020

Haemin pre-treatment augments the cardiac protection of mesenchymal stem cells by inhibiting mitochondrial fission and improving survival.

Journal of cellular and molecular medicine

Deng R,Liu Y,He H,Zhang H,Zhao C,Cui Z,Hong Y,Li X,Lin F,Yuan D,Liang X,Zhang Y

PA5-64821 was used in Western Blotting to study regulation of mesenchymal stem cell survival by haemin.

Wed Jan 01 00:00:00 EST 2020

Vascular progenitor cell senescence in patients with Marfan syndrome.

Journal of cellular and molecular medicine

He H,Yu B,Liu Z,Ye G,You W,Hong Y,Lian Q,Zhang Y,Li X

PA5-64821 was used in Western Blotting to assess the function of Vascular progenitor cells (VPCs) isolated from patients with Marfan syndrome (MFS).

Sat Jun 01 00:00:00 EDT 2019

Cardiac Dysfunction in the Sigma 1 Receptor Knockout Mouse Associated With Impaired Mitochondrial Dynamics and Bioenergetics.

Journal of the American Heart Association

Abdullah CS,Alam S,Aishwarya R,Miriyala S,Panchatcharam M,Bhuiyan MAN,Peretik JM,Orr AW,James J,Osinska H,Robbins J,Lorenz JN,Bhuiyan MS

PA5-64821 was used in Western Blotting to investigate the physiological function of Sigmar1 in regulating cardiac hemodynamics using the Sigmar1 knockout mouse (Sigmar1-/-).

Tue Oct 16 00:00:00 EDT 2018

Bioinformatics

Protein Aliases: C-terminal region; Dnm1p/Vps1p-like protein; DVLP; Dymple; dynamin 1-like; Dynamin family member proline-rich carboxyl-terminal domain less; Dynamin-1-like protein; Dynamin-like protein; dynamin-like protein 1; Dynamin-like protein 4; Dynamin-like protein IV; Dynamin-related protein 1; DYNIV 11; DYNIV-11; EC 3.6.5.5; FLJ41912; HdynIV; N-terminal region; VPS1

Gene Aliases: 6330417M19Rik; AI450666; DLP1; DNM1L; Dnml1; Dnmlp1; DRP1; DVLP; DYMPLE; EMPF; HDYNIV; python

UniProt ID: (Human) O00429, (Rat) O35303, (Mouse) Q8K1M6

Entrez Gene ID: (Human) 10059, (Rat) 114114, (Mouse) 74006

Molecular Function: membrane traffic protein

mitochondrial fission release of cytochrome c from mitochondria dynamin polymerization involved in mitochondrial fission endocytosis cellular component disassembly involved in execution phase of apoptosis regulation of mitochondrion organization peroxisome fission regulation of protein oligomerization positive regulation of apoptotic process positive regulation of GTPase activity mitochondrial fragmentation involved in apoptotic process positive regulation of protein secretion protein oligomerization protein homotetramerization heart contraction membrane fusion necroptotic process mitochondrion morphogenesis protein localization to mitochondrion positive regulation of mitochondrial fission mitochondrial membrane fission positive regulation of release of cytochrome c from mitochondria regulation of peroxisome organization regulation of mitophagy regulation of ATP metabolic process positive regulation of intrinsic apoptotic signaling pathway metabolic process programmed cell death

Process(es)

GTPase activity GTPase activator activity protein binding GTP binding lipid binding Rab GTPase binding GTP-dependent protein binding ubiquitin protein ligase binding protein homodimerization activity identical protein binding nucleotide binding hydrolase activity