MA5-24859

antibody from Invitrogen Antibodies

Targeting: TOMM20 KIAA0016, MAS20, MOM19, TOM20

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-24859

Provider

Invitrogen Antibodies

Product name

TOMM20 Recombinant Rabbit Monoclonal Antibody (3G5)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Antibody clone number

3G5

Vial size

100 µL

Concentration

1 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 TOMM20 in Lane 1: HeLa whole cell lysate, Lane 2: MCF7 whole cell lysate, Lane 3: F9 whole cell lysate, Lane 4: PC12 whole cell lysate. Samples were incubated with TOMM20 monoclonal antibody (Product # MA5-24859) at a dilution of 1:1,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of TOMM20 in Lane 1: HeLa whole cell lysate, Lane 2: MCF7 whole cell lysate, Lane 3: F9 whole cell lysate, Lane 4: PC12 whole cell lysate. Samples were incubated with TOMM20 monoclonal antibody (Product # MA5-24859) at a dilution of 1:1,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of TOMM20 was achieved by transfecting HeLa cells with TOMM20 specific siRNAs (Silencer® select Product # s18949, s18948). Western blot analysis (Fig. a) was performed using membrane extracts from the HeLa knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with TOMM20 Recombinant Rabbit Monoclonal Antibody (3G5) (Product # MA5-24859, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (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 TOMM20.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-TOMM20 Recombinant Rabbit Monoclonal Antibody (3G5) (Product # MA5-24859) and a 15kDa band corresponding to TOMM20 was observed across all the cell lines and tissue tested. Membrane extracts (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), MCF-7 (Lane 3), SH-SY5Y (Lane 4), THP-1 (Lane 5), HL-60 (Lane 6), tissue extracts of Mouse Brain (Lane 7), Mouse Kidney (Lane 8) and Rat Brain (Lane 9) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of TOMM20 in PC12 cells (green). Samples were treated with paraformaldehyde, permeabilized with 0.25% Triton and PBS, and incubated with TOMM20 monoclonal antibody (Product # MA5-24859).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of TOMM20 in HepG2 cells (green). Samples were treated with paraformaldehyde, permeabilized with 0.25% Triton and PBS, and incubated with TOMM20 monoclonal antibody (Product # MA5-24859).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis was performed using Anti-TOMM20 Rabbit Monoclonal Antibody (Product # MA5-24859) using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with TOMM20 Rabbit Monoclonal Antibody (Product # MA5-24859) at 1:100 dilution in 0.1% BSA and incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing mitochondrial localization. Panel e represents the no primary control. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of TOMM20 in paraffin-embedded mouse small intestine tissue. Samples were incubated with TOMM20 monoclonal antibody (Product # MA5-24859) followed by hematoxylin.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of TOMM20 in paraffin-embedded mouse kidney tissue. Samples were incubated with TOMM20 monoclonal antibody (Product # MA5-24859) followed by hematoxylin.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of TOMM20 in paraffin-embedded mouse heart tissue. Samples were incubated with TOMM20 monoclonal antibody (Product # MA5-24859) followed by hematoxylin.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of TOMM20 in paraffin-embedded human liver cancer tissue. Samples were incubated with TOMM20 monoclonal antibody (Product # MA5-24859) followed by hematoxylin.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of TOMM20 in paraffin-embedded human kidney tissue. Samples were incubated with TOMM20 monoclonal antibody (Product # MA5-24859) followed by hematoxylin.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of TOMM20 in HeLa cells (blue) and compared with an unlabeled control (cells without incubation with primary antibody; red). Samples were incubated with TOMM20 monoclonal antibody (Product # MA5-24859) at a dilution of 1:50 followed by Alexa Fluor 488-conjugated goat anti rabbit IgG.