Invitrogen

Dynein Monoclonal Antibody (74.1)

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

Dynein Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Dynein in U87-MG Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a Dynein monoclonal antibody (Product # MA1-070) at a dilution of 1:20 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). Dynein staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

Dynein Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of Dynein in U87-MG cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant, adding 90% methanol and incubated for 10 minutes at room temperature. Follwing penetration, cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a Dynein monoclonal antibody (Product # MA1-070) at a dilution of 2 µg/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

Dynein Antibody in Immunoprecipitation (IP) — Immunoprecipitation of Dynein was performed on K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 µg of Dynein monoclonal antibody (Product # MA1-070) rotating 60 min at RT. The immune complexes were captured on 625 µg of anti-mouse coated Dynabeads (Product # 11202D), washed extensively, and eluted with NuPAGE™ LDS Sample Buffer (Product # NP0007). Samples were resolved onto NuPAGE™ 4-12% Bis-Tris gel (Product # NP0335BOX). Lanes 1 and 3 are input and lanes 2 and 4 are IP. Proteins were transferred to PVDF membrane (Product # IB23001). Membrane was blocked in 5% milk. Target was detected using a Dynein monoclonal antibody (Product # MA1-070) at a dilution of 1:2000, followed by a 1:4000 dilution of secondary antibody. Chemiluminescent detection was performed using ECL Western Blotting Substrate (Product # 32106). Data courtesy of the Yeo lab as part of the ENCODE project (www.encodeproject.org).

Dynein Antibody in RNA Immunoprecipitation (RIP) — RNA immunoprecipitation (RIP) western of Dynein was performed on K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 µg of Dynein monoclonal antibody (Product # MA1-070) rotating 60 min at RT. The immune complexes were captured on 625 µg of anti-mouse coated Dynabeads (Product # 11202D), washed extensively, and eluted with NuPAGE™ LDS Sample Buffer (Product # NP0007). Samples were resolved onto NuPAGE™ 4-12% Bis-Tris gel (Product # NP0335BOX). Lanes 1 and 3 are input and lanes 2 and 4 are IP. Proteins were transferred to PVDF membrane (Product # IB23001). Membrane was blocked in 5% milk. Target was detected using a Dynein monoclonal antibody (Product # MA1-070) at a dilution of 1:2000, followed by a 1:4000 dilution of secondary antibody. Chemiluminescent detection was performed using ECL Western Blotting Substrate (Product # 32106). Data courtesy of the Yeo lab as part of the ENCODE project (www.encodeproject.org).

Product Details

MA1-070

Applications

Tested Dilution

Publications

Western Blot (WB)

1 µg/mL

View 4 publications 4 publications

Immunocytochemistry (ICC/IF)

2-4 µg/mL

View 1 publication 1 publication

Flow Cytometry (Flow)

1-2 µg/test

View 1 publication 1 publication

Immunoprecipitation (IP)

Assay-dependent

View 2 publications 2 publications

RNA Immunoprecipitation (RIP)

Assay-dependent

Product Specifications

Species Reactivity

Bovine, Human, Many, Mouse, Rat

Published species

Human, Mouse, Rat

Host/Isotype

Mouse / IgG2b

Class

Monoclonal

Type

Antibody

Clone

74.1

Immunogen

Purified bovine brain cytoplasmic dynein.

View immunogen

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

PBS with 1mg/mL BSA

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_2093668

Product Specific Information

MA1-070 detects two isoforms of the ~74 kDa polypetide of cytoplasmic dynein.

MA1-070 has been successfully used in Western blot immunofluorescence, and immunoprecipitation procedures. In Western blot analysis of HeLa cell lysate this antibody detects a ~74 kDa protein representing cytoplasmic dynein.

The MA1-070 immunogen is purified bovine brain cytoplasmic dynein.

Target Information

The directed movement of membrane-bounded organelles is an important process for various cellular functions including membrane transport, secretion, and axonal transport. Cytoplasmic dynein, a ubiquitous minus end-directed microtubule-based motor protein, is believed to be responsible for the retrograde transport of membranous organelles from the synapse to the cell body in fast axonal transport. The cytoplasmic dynein complex is composed of two ~530 kDa heavy chains, and intermediate chains of ~74 kDa and ~53-59 kDa.

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

Product References

Cilia proteins are biomarkers of altered flow in the vasculature.

JCI insight

Gupta A,Thirugnanam K,Thamilarasan M,Mohieldin AM,Zedan HT,Prabhudesai S,Griffin MR,Spearman AD,Pan A,Palecek SP,Yalcin HC,Nauli SM,Rarick KR,Zennadi R,Ramchandran R

MA1-070 was used in Flow Cytometry to suggest that cilia and ciliary proteins in circulation are detectable under various altered-flow conditions, which could serve as a surrogate biomarker of the damaged endothelium.

Tue Mar 22 00:00:00 EDT 2022

Sorting Nexin 27 Enables MTOC and Secretory Machinery Translocation to the Immune Synapse.

Frontiers in immunology

González-Mancha N,Rodríguez-Rodríguez C,Alcover A,Merida I

MA1-070 was used in Western Blot, Immunocytochemistry to show that sorting nexin 27 modulates immune synapse organisation through regulated trafficking of cargoes in T lymphocytes.

Mon Mar 07 00:00:00 EST 2022

Sorting Nexin 27 Enables MTOC and Secretory Machinery Translocation to the Immune Synapse.

Frontiers in immunology

González-Mancha N,Rodríguez-Rodríguez C,Alcover A,Merida I

MA1-070 was used in Western Blot, Immunocytochemistry to show that sorting nexin 27 modulates immune synapse organisation through regulated trafficking of cargoes in T lymphocytes.

Mon Mar 07 00:00:00 EST 2022

KIF5A-dependent axonal transport deficiency disrupts autophagic flux in trimethyltin chloride-induced neurotoxicity.

Autophagy

Liu M,Pi H,Xi Y,Wang L,Tian L,Chen M,Xie J,Deng P,Zhang T,Zhou C,Liang Y,Zhang L,He M,Lu Y,Chen C,Yu Z,Zhou Z

MA1-070 was used in Western Blot to investigate the mechanism of induction of neurotoxicity by trimethyltin chloride.

Thu Apr 01 00:00:00 EDT 2021

Two new isoforms of the human hepatoma-derived growth factor interact with components of the cytoskeleton.

Biological chemistry

Nüße J,Mirastschijski U,Waespy M,Oetjen J,Brandes N,Rebello O,Paroni F,Kelm S,Dietz F

MA1-070 was used in western blot to study the interaction of components of the cytoskeleton with two new isoforms of the human hepatoma-derived growth factor

Sun May 01 00:00:00 EDT 2016

Cytoplasmic dynein regulates the subcellular distribution of mitochondria by controlling the recruitment of the fission factor dynamin-related protein-1.

Journal of cell science

Varadi A,Johnson-Cadwell LI,Cirulli V,Yoon Y,Allan VJ,Rutter GA

MA1-070 was used in immunoprecipitation to investigate the regulation of cellular localization of mitochondria by dynein

Wed Sep 01 00:00:00 EDT 2004

Differential expression and phosphorylation of the 74-kDa intermediate chains of cytoplasmic dynein in cultured neurons and glia.

The Journal of biological chemistry

Pfister KK,Salata MW,Dillman JF,Vaughan KT,Vallee RB,Torre E,Lye RJ

MA1-070 was used in western blot to investigate the brain cytoplasmic IC74 isoforms and their interaction with dynein

Fri Jan 19 00:00:00 EST 1996

Cytoplasmic dynein is associated with slow axonal transport.

Proceedings of the National Academy of Sciences of the United States of America

Dillman JF,Dabney LP,Pfister KK

MA1-070 was used in immunoprecipitation to investigate the relationship between dynein and slow axonal transport

Tue Jan 09 00:00:00 EST 1996

Bioinformatics

Protein Aliases: 8 kDa dynein light chain; 8 kDa dynein light chain b; 8kD LC; 8kDa LC; cytoplasmic dynein light polypeptide; cytosolic DH IC-1; DLC8; DLC8b; DNCL1; dynein LC8; Dynein light chain 1, cytoplasmic; Dynein light chain 2, cytoplasmic; Dynein light chain LC8-type 1; Dynein light chain LC8-type 2; dynein light chain-2; dynein, cytoplasmic, light chain 1; dynein, cytoplasmic, light peptide; dynein, cytoplasmic, light polypeptide 1; dynein, light chain, LC8-type 1; dynein, light chain, LC8-type 2; mPIN; PIN; Protein inhibitor of neuronal nitric oxide synthase; protein inhibitor of nitric oxide synthase; radial spoke 22 homolog

Gene Aliases: 1700064A15Rik; 6720463E02Rik; 8kDLC; C87222; CDLC2; DLC1; DLC2; DLC8; DLC8b; DNCL1; DNCL1B; DNCLC1; DYNLL1; DYNLL2; HDLC1; LC8; LC8a; PIN; RSPH22

UniProt ID: (Human) P63167, (Bovine) P61285, (Human) Q96FJ2, (Bovine) Q3MHR3, (Rat) P63170, (Mouse) P63168, (Rat) Q78P75, (Mouse) Q9D0M5

Entrez Gene ID: (Human) 8655, (Bovine) 404151, (Human) 140735, (Bovine) 540369, (Rat) 58945, (Mouse) 56455, (Rat) 140734, (Mouse) 68097

Molecular Function: microtubule or microtubule-binding cytoskeletal protein