MGMT Rabbit Monoclonal Antibody(ARA735)

Datasheet

MSDS

Key features and details

Target: MGMT
Source/Host: Rabbit
Reactivity: Human
Clonality: Monoclonal
Applications: WB,IHC,FC,IP
Conjugation: Unconjugated
Storage: at-20°C

Brand:

CAT.NO. : ARA6522

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Size:

100μL

Trail, Bulk size or Custom requests Please contact us

Product Details

Background

Involved in the cellular defense against the biological effects of O6-methylguanine (O6-MeG) and O4-methylthymine (O4-MeT) in DNA. Repairs the methylated nucleobase in DNA by stoichiometrically transferring the methyl group to a cysteine residue in the enzyme. This is a suicide reaction: the enzyme is irreversibly inactivated.

Application

To ensure optimal assay performance, AREX recommends conducting reagent titration tailored to each testing system for optimal detection results.

WB	1:5000-1:10000
IHC	1:100-1:200
FC	1:10-1:200
IP	1:80

*Results are sample-specific. Please refer to your local assay conditions and test parameters for reference.

Overview

Description	Rabbit Monoclonal Antibody to MGMT
Antibody Type	Primary antibody
Predicted MW	22kDa
Immunogen	A synthetic peptide corresponding to residues on the N-terminus of human MGMT was used as an immunogen.
Purification	ProA affinity purified IgG
Form/Buffer	PBS 59%, Sodium azide 0.01%, Glycerol 40%, BSA 0.10%.
Alternative Names	Methylated-DNA--protein-cysteine methyltransferase; 6-O-methylguanine-DNA methyltransferase; MGMT; O-6-methylguanine-DNA-alkyltransferase
Gene Symbol	MGMT
Entrez Gene	4255(Human)
Swissprot	P16455

*AREX continuously optimizes our products. Webpage content may not reflect the latest updates. For inquiries, please contact info@arexbio.com or your local distributor.
*Clone Number, Reactivity, Source/Host and Clonality can be found in the product name and Key Features section above.

Data

All lanes: Anti-MGMT antibody
at 1:5,000 dilution
Predicted MW: 22 kDa
Observed MW: 22 kDa

Lane 1: MCF-7
Lane 2: Jurkat
Lane 3: Hela
Lane 4: HT-29

Lysates at 20 µg per lane
2nd Ab:
GAR HRP(H+L) 1:10,000
Exposure: 120s

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast tissue labelling MGMT with ARA735 at 1:200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0.

Overlay histogram showing Jurkat cells stained with ARA735 (Blue). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% TritonX-100 for 15 min. The cells were then incubated in the antibody (ARA735, 1:50 dilution) in 1x PBS/1% BSA for 30 min at 4°C. The secondary antibody used was a Goat Anti-Rabbit Alexa Fluor<sup>®</sup> 488 (IgG H+L) at 1:2,000 dilution for 20 min at 4°C.
Unlabelled sample (Red) was used as a control.

MGMT was immunoprecipitated from 0.4 mg of Hela whole cell lysate with ARA735 at 1:80 dilution.
2nd Ab:
GAR HRP for IP 1:500

Lane 1: ARA735 IP in Hela whole cell lysate
Lane 2: PBS instead of ARA735 in Hela whole cell lysate
Lane 3: Hela whole cell lysate, 10 µg (input)

Exposure: 120s

Storage

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

Note

For Research Use Only. Not for diagnostic, therapeutics, prophylactic or in vivo use.

FAQs

What are the main types of research antibodies and how do they differ?

Research antibodies are mainly divided into monoclonal antibodies and polyclonal antibodies. Monoclonal antibodies typically offer higher specificity and better batch-to-batch consistency, while polyclonal antibodies often provide stronger affinity but may show more variation between batches. The choice depends on your specific experimental needs.

How can I tell if a research antibody is suitable for my experiment?

It is recommended to carefully review the product datasheet for validated applications, species reactivity, recommended dilutions, and published references. For new antibodies, performing a small-scale validation with positive control samples is usually helpful.

Can improper storage of research antibodies affect experimental results?

Yes. Antibodies are sensitive to temperature, repeated freeze-thaw cycles, and contamination. Improper storage may lead to reduced activity, increased background, or weaker signals. It is best to follow the storage instructions provided in the product datasheet.

Why doesn’t the recommended dilution in the datasheet work well in my experiment?

The recommended dilution is based on the supplier’s test conditions. Factors such as sample type, fixation method, and detection system in your lab can influence the optimal working concentration. Performing a dilution series optimization in your own system is often necessary.

What precautions should I take when using a newly purchased research antibody for the first time?

It is advisable to briefly centrifuge the antibody (especially concentrated or lyophilized ones), then perform a small-scale pilot experiment using the recommended conditions. Recording the batch number and usage date is also helpful for future tracking.

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