Invitrogen

TOP1 Recombinant Rabbit Monoclonal Antibody (SC69-03)

Advanced Verification

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

1 Published Figure

1 Reference

View all (24) Topoisomerase I antibodies

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FIGURE: 1 / 12

TOP1 Antibody (MA5-32228) in WB

Western blot was performed using Anti-TOP1 Recombinant Rabbit Monoclonal Antibody (SC69-03) (Product # MA5-32228) and a 91 kDa band corresponding to TOP1 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), MCF-7 (Lane 2), Hep G2 (Lane 3), Jurkat (Lane 4), NTERA-2 cl.D1 (Lane 5) and NIH/3T3 (Lane 6) were electrophoresed using 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 S... View More

Product Image Gallery

TOP1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of TOP1 was performed using 70% confluent log phase HeLa 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 TOP1 Recombinant Rabbit Monoclonal Antibody (SC69-03) (Product # MA5-32228) at 5 µg/mL in 0.1% BSA, 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). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

TOP1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of Topoisomerase I of paraffin-embedded Mouse brain tissue using a Topoisomerase-I Monoclonal antibody (Product #MA5-32228). Counter stained with hematoxylin.

TOP1 Antibody in Flow Cytometry (Flow) — Flow Cytometric analysis of Topoisomerase I in HepG2 cells using a Topoisomerase I Monoclonal Antibody (Product # MA5-32228) at a dilution of 1:50, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.

TOP1 Antibody in ChIP Assay (ChIP) — Chromatin Immunoprecipitation (ChIP) assay of endogenous TOP1 protein using Anti-TOP1 Antibody: ChIP was performed using Anti-TOP1 Rabbit Recombinant Monoclonal Antibody (Product # MA5-32228, 5 µg) on sheared chromatin from Camptothecin treated HCT 116 cells (10uM, 2hours) (using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to CDKN1A intron 1, 18s rDNA, promoter and transcriptional start site of GAPDH, and SAT2 satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

TOP1 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with TOP1 siRNA and decrease in signal intensity was observed in western blot application using TOP1 Recombinant Rabbit Monoclonal Antibody (SC69-03) (Product # MA5-32228). {KD}

Product Details

MA5-32228

Applications

Tested Dilution

Publications

Western Blot (WB)

1:1,000-1:5,000

View 1 publication 1 publication

Immunohistochemistry (IHC)

1:50-1:1,000

Immunocytochemistry (ICC/IF)

1:50-1:500

Flow Cytometry (Flow)

1:50-1:100

ChIP assay (ChIP)

2.5 µg/10^6 cells

Product Specifications

Species Reactivity

Human, Mouse

Published species

Human, Mouse

Host/Isotype

Rabbit / IgG

Expression System

HEK293 cells

Class

Recombinant Monoclonal

Type

Antibody

Clone

SC69-03

Immunogen

Synthetic peptide within Human TOP1 aa 1-49

View immunogen

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

TBS, pH 7.4, with 40% Glycerol, 0.05% BSA

Contains

0.05% sodium azide

Storage conditions

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

RRID

AB_2809514

Product Specific Information

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.

Target Information

This gene encodes a DNA topoisomerase, an enzyme that controls and alters the topologic states of DNA during transcription. This enzyme catalyzes the transient breaking and rejoining of a single strand of DNA which allows the strands to pass through one another, thus altering the topology of DNA. This gene is localized to chromosome 20 and has pseudogenes which reside on chromosomes 1 and 22.

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

Product References

A brain proteomic signature of incipient Alzheimer's disease in young APOE ε4 carriers identifies novel drug targets.

Science advances

Roberts JA,Varma VR,An Y,Varma S,Candia J,Fantoni G,Tiwari V,Anerillas C,Williamson A,Saito A,Loeffler T,Schilcher I,Moaddel R,Khadeer M,Lovett J,Tanaka T,Pletnikova O,Troncoso JC,Bennett DA,Albert MS,Yu K,Niu M,Haroutunian V,Zhang B,Peng J,Croteau DL,Resnick SM,Gorospe M,Bohr VA,Ferrucci L,Thambisetty M

MA5-32228 was used in Western Blot to characterize and validate the precise mechanisms mediating APOE ε4-associated risk of Alzheimer’s disease (AD), and show that drugs targeting three of these proteins—STAT3, YES1, and FYN—rescue distinct molecular phenotypes relevant to AD pathogenesis.