702782

antibody from Invitrogen Antibodies

Targeting: DNM1L DRP1, DVLP, DYMPLE, HDYNIV, VPS1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

702782

Provider

Invitrogen Antibodies

Product name

DNM1L Recombinant Rabbit Monoclonal Antibody (2H15L40)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Antibody clone number

2H15L40

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of DNM1 was performed by loading 30µg of MCF7 CAS9 Control (lane1), MC7- DNM1L knockout (lane2) whole cell lysate using Novex® NuPAGE® 4-12 % Bis-Tris gel (NP0321BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard  (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk in PBST for 1 hour at room temperature. DNM1L was detected at ~80 kDa using Anti-DNM1L Rabbit Monoclonal Antibody (702782) at 1:250 dilution in 5% skim milk PBST at 4°C overnight on a gentle rocking platform. Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to DNM1L.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Whole cell extracts (30 µg lysate) of SH-SY5Y (Lane 1), NTERA-2 (Lane 2) and tissue extracts of Mouse Brain (Lane 3), Rat Brain (Lane 4), Mouse Heart (Lane 5) and Rat Heart (Lane 6). The blots were probed with Anti-DNM1L Recombinant Rabbit Monoclonal Antibody (Product # 702782, 1:200 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036,1:4000 dilution). A~ 80 kDa band corresponding to DNM1L was observed across the cell lines and tissues tested.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, SH-SY5Y cells were fixed and permeabilized for detection of endogenous DNM1L using Anti- DNM1L Recombinant Rabbit Monoclonal Antibody (Product # 702782, 1:100 dilution) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000 dilution). Panel a) shows representative cells that were stained for detection and localization of DNM1L protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of DNM1L. Panel e) represents control cells without primary antibody to assess background. The images were captured at 60X magnification.