- MA5-26255 - Invitrogen Antibodies DNM1L antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of DNM1L in human tissue (1: Uterus, 2: Breast, 3: Brain, 4: Liver, 5: Ovary, 6: Thyroid, 7: colon) tissue using 10 µg per lane. Samples were separated by SDS-PAGE and probed with DNM1L (Product # MA5-26255) monoclonal antibody at a dilution of 1:200.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of DNM1L in HEK293T cells in untransfected (Left lane) and transfected (Right lane) samples using 5 µg per lane. The samples were separated by SDS-PAGE and probed with DNM1L (Product # MA5-26255) monoclonal antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of DNM1L in HepG2, HeLa, SVT2, A549, COS7, Jurkat, MDCK, PC12 and MCF7 cells using 35 µg per lane. Samples were separated by SDS-PAGE and probed with DNM1L (Product # MA5-26255) monoclonal antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of DNM1L was achieved by transfecting SH-SY5Y with DNM1L specific siRNAs (Silencer® select Product # S19559, S19561). Western blot analysis (Fig. a) was performed using Whole cell extracts from the DNM1L knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with DNM1L Monoclonal Antibody (OTI4F6) (Product # MA5-26255, 1:2000 dilution ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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 DNM1L.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-DNM1L Monoclonal Antibody (OTI4F6)(Product # MA5-26255) and a 80kDa band corresponding to DNM1L was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of SH-SY5Y (Lane 1), NTERA-2 cl.D1 (Lane 2), SK-O-V3 (Lane 3), PC-12 (Lane 4), Mouse Brain (Lane 5) and Rat Brain (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 (1:2000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of DNM1L was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with DNM1L Monoclonal Antibody (OTI4F6) (Product # MA5-26255) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing mitochondria and peroxisome like cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded human pancreas tissue. To expose target proteins, 10mM citric buffer, pH6.0, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a DNM1L monoclonal antibody (Product # MA5-26255).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded carcinoma of human lung tissue. To expose target proteins, 10mM citric buffer, pH6.0, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a DNM1L monoclonal antibody (Product # MA5-26255).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded carcinoma of human pancreas tissue. To expose target proteins, 10mM citric buffer, pH6.0, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a DNM1L monoclonal antibody (Product # MA5-26255).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded adenocarcinoma of human ovary tissue. To expose target proteins, 10mM citric buffer, pH6.0, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a DNM1L monoclonal antibody (Product # MA5-26255).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded human endometrium tissue. To expose target proteins, 10mM citric buffer, pH6.0, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a DNM1L monoclonal antibody (Product # MA5-26255).

MA5-26255