GENTAUR Belgium BVBA BE0473327336 Voortstraat 49, 1910 Kampenhout BELGIUM Tel 0032 16 58 90 45
GENTAUR U.S.A Genprice Inc 6017 Snell Ave, Ste 357, SanJose, CA 95123
Tel (408) 780-0908, Fax (408) 780-0908, sales@genprice.com

Index / EpigenTek / EpiQuik MBD2 Binding Activity_Inhibition Assay Kit / Product Detail : P-3021-48 EpiQuik MBD2 Binding Activity_Inhibition Assay Kit
Related keywords:

Aug
23th

#P-3021-48 EpiQuik MBD2 Binding Activity_Inhibition Assay Kit

Ask technical file .

  Price : 462   EUR
524   USD
358   GBP
1941   Zloty
61836   JPY
3564   NOK
3818   SEK
522   CHF

Product name : EpiQuik MBD2 Binding Activity_Inhibition Assay Kit

Catalog number : P-3021-48

Quantity: 48 assays

Availability: Yes

Supplier name : EpigenTek

ask pdf gentaur products Data sheet: Ask more or other datasheet now !

More Details about


EpiQuik™ MBD2 Binding Activity Assay Kit
Catalog No. P-3021


INTRODUCTION


MBD2 (methyl-CpG-binding domain protein 2) is a member of the MBD protein family. MBD2 selectively binds to methylated DNA and suppresses transcription from a methylated target gene through recruiting transcriptional repressor complexes, which contain Mi-2/NuRD or HDACs. MBD2 has also been shown to catalyze demethylation by directly removing methyl groups from 5-methylcytosine residues in DNA. MBD2 is demonstrated to be associated with tumorigenesis. For example, deficiency of MBD2 suppresses intestinal tumor formation, indicating that MBD2 is not necessary only for tumor development, but also for tumor growth. The binding activity of MBD2 to methylated DNA may be affected by MBD2 mutation and by biochemical or pharmacological intervention. So far, there are few assays available for measuring MBD2 binding activity in vitro. The EpiQuik™ MBD2 Binding Activity Assay Kit provides a unique procedure to measure binding activity of MBD2 to methylated DNA. The kit has following features:


• The fastest procedure, which can be finished within 3 hours.
• Innovative colorimetric assay to quantitatively measure MBD2 binding activity.
• Strip microplate format makes the assay flexible: manual or high throughput analysis.
• Simple, reliable, and consistent assay conditions.


PRINCIPLE AND PROCEDURE


The EpiQuik™ MBD2 Binding Activity Assay Kit is designed for measuring in vitro or intracellular MBD2 binding activity. In an assay with this kit, activated MBD2 protein contained in nuclear extract binds to methylated DNA and forms the MBD2/methylated DNA complex. The complex is then captured to a specificallytreated strip well plate. MBD2 is recognized with a specific antibody and measured colorimetrically.

PRODUCT USE INFORMATION


The EpiQuik™ MBD2 Binding Activity Assay Kit is suitable for measuring MBD2 binding activity in human cells/tissues.
Epigentek guarantees the performance of all products in the manner described in our product instructions.
Epigentek reserves the right to change or modify any product to enhance its performance and design.
The EpiQuik™ MBD2 Binding Activity Assay Kit is for research use only and is not intended for diagnostic or therapeutic application.
EpiQuik™ is a trademark of Epigentek Group Inc.
The EpiQuik™ MBD2 Binding Activity Assay Kit and method of use are covered by a pending US patent.


KIT CONTENTS


Components                                                 48 assays                96 assays
                                                                         P-3021-48                P-3021-96
MB1 (10X Wash Buffer)                               12 ml                           25 ml
MB2 (Assay Buffer)                                      1.5 ml                             3 ml
MB3 (Methylated DNA, 25 μg/ml)*               50 μl                         100 μl
MB4 (Affinity Antibody, 100 μg/ml)*              25 μl                           50 μl
MB5 (Detection Antibody, 200 μg/ml)*        10 μl                           20 μl
MB6 (Developing Solution)                             6 ml                         12 ml
MB7 (Stop Solution)                                         3 ml                           6 ml
8-Well Assay Strips (with Frame)                  6                                 12

* For maximum recovery of the products, centrifuge the original vial after thawing prior toopening the cap.


SHIPPING AND STORAGE
The kit is shipped in two parts: one part at ambient room temperature, and the second part on frozen ice packs at 4°C. Upon receipt: (1) Store MB5 at –20°C away from light; (2) Store MB1, MB3, MB4, MB6 and 8-Well Assay Strips at 4°C away from light; and (3) Store all other components (MB2, MB7) at room temperature. The kit is stable for up to 6 months from the shipment date, when stored properly.

Note: Check if wash buffer, MB1, contains salt precipitates before using. If so, warm (at room temperature or 37°C) and shake the buffer until the salts are redissolved.


MATERIALS REQUIRED BUT NOT SUPPLIED
Orbital shaker
Pipettes and pipette tips
Microplate reader
1.5 ml microcentrifuge tubes


PROTOCOL
1. Prepare nuclear extracts from treated and untreated cells or tissues by using you own successful method. For your convenience and the best results, Epigentek offers the EpiQuik™ Nuclear Extraction Kit (Cat. No. OP-0002-1) optimized for use in the EpiQuik™ series. Nuclear extracts can be used immediately or stored at -80°C for future use.


2. Determine the number of strip wells required. Leave these strips in the plate frame (remaining unused strips can be placed back in the bag. Seal the bag tightly and store at 4°C). Dilute with distilled water (pH 7.2 to 7.5) at a 1:10 ratio (e.g., 1 ml of GU1 + 9 ml of distilled water). Wash the strip wells once with 150 μl of the diluted MB1.


3. Add 26 μl of MB2, 1 μl of MB3, and then add 3 μl of nuclear extracts (4-20 μg) or purified MBD2 protein to each strip well. Mix and cover the strip wells with Parafilm M and incubate at 37°C for 60 minutes. For the blank, add 3 μl of MB2 instead of nuclear extracts.


4. Aspirate and wash each well with 150 μl of diluted MB1 three times.


5. Dilute MB4 (at a 1:100 ratio) to 1 μg/ml with diluted MB1. Add 50 μl of the diluted MB4 to each strip well and incubate at room temperature for 60 minutes on an orbital shaker (50-100 rpm).
6. Aspirate and wash each well with 150 μl of diluted MB1 four times.

7. Dilute MB5 (at a 1:1000 ratio) to 1 μg/ml with diluted MB1. Add 50 μl of the diluted MB5 to each strip well and incubate at room temperature for 30 minutes.

8. Aspirate and wash each well with 150 μl of diluted MB1 four times. Allow 3 minutes for last wash.

9. Add 100 μl of MB6 to each well and incubate at room temperature for 2-10 minutes away from light. Monitor the color development in the sample and control wells (blue).

10. Add 50 μl of MB7 to each well to stop enzyme reaction when the color in the standard wells containing the higher concentrations of standard control turns medium blue. The color should change to yellow and absorbance can be read on a microplate reader at 450 nm within 2-15 min

11. Calculate % binding of MBD2:

TROUBLESHOOTING
No Signal for the Sample
High Background Present for the Blank

Contact us about this product :

Our team will respond you as soon as possible !

Email :
Skype :
Name :
Phone :
address :
Question, Comment ... :
arrow security gentaurPlease retype this code below:
EpigenTek \ EpiQuik_MBD2_Binding_Activity_Inhibition_Assay_Kit___ \ P_3021_48
Reload Image

 

GENTAUR Belgium BVBA BE0473327336
Voortstraat 49, 1910 Kampenhout BELGIUM
Tel 0032 16 58 90 45

Fax 0032 16 50 90 45
info@gentaur.com | Gentaur





GENTAUR Ltd.
Howard Frank Turnberry House
1404-1410 High Road
Whetstone London N20 9BH
Tel 020 3393 8531 Fax 020 8445 9411
uk@gentaur.com | Gentaur

 

 




GENTAUR France SARL
9, rue Lagrange, 75005 Paris
Tel 01 43 25 01 50

Fax 01 43 25 01 60
RCS Paris B 484 237 888

SIRET 48423788800017

BNP PARIBAS PARIS PL MAUBERT BIC BNPAFRPPPRG

france@gentaur.com | Gentaur

GENTAUR GmbH
Marienbongard 20
52062 Aachen Deutschland
Support Karolina Elandt
Tel: 0035929830070
Fax: (+49) 241 56 00 47 88

Logistic :0241 40 08 90 86
Bankleitzahl 39050000
IBAN lautet DE8839050000107569353
Handelsregister Aachen HR B 16058
Umsatzsteuer-Identifikationsnummer *** DE 815175831
Steuernummer 201/5961/3925
de@gentaur.com | Gentaur

GENTAUR U.S.A
Genprice Inc, Logistics
547, Yurok Circle
San Jose, CA 95123
CA 95123
Tel (408) 780-0908,
Fax (408) 780-0908,
sales@genprice.com

Genprice Inc, Invoices and accounting
6017 Snell Ave, Ste 357
San Jose, CA 95123




GENTAUR Nederland BV
NL850396268B01 KVK nummer 52327027
Kuiper 1
5521 DG Eersel Nederland
Tel:  0208-080893  Fax: 0497-517897
nl@gentaur.com | Gentaur
IBAN: NL04 RABO 0156 9854 62   SWIFT RABONL2U






GENTAUR Spain
tel:0911876558
spain@gentaur.com | Gentaur






ГЕНТАУЪР БЪЛГАРИЯ
ID # 201 358 931 /BULSTAT
София 1000, ул. "Граф Игнатиев" 53 вх. В, ет. 2
Tel 0035924682280 Fax 0035924808322
e-mail: Sofia@gentaur.com | Gentaur
IBAN: BG11FINV91501014771636
BIC: FINVBGSF

GENTAUR Poland Sp. z o.o.


ul. Grunwaldzka 88/A m.2
81-771 Sopot, Poland
TEL Gdansk 058 710 33 44 FAX  058 710 33 48              

poland@gentaur.com | Gentaur

Other countries

Österreich +43720880899

Canada Montreal +15149077481

Ceská republika Praha +420246019719

Danmark +4569918806

Finland Helsset +358942419041

Magyarország Budapest +3619980547

Ireland Dublin+35316526556

Luxembourg+35220880274

Norge Oslo+4721031366

Sverige Stockholm+46852503438

Schweiz Züri+41435006251

US New York+17185132983

GENTAUR Italy
SRL IVA IT03841300167
Piazza Giacomo Matteotti, 6
24122 Bergamo Tel 02 36 00 65 93
Fax 02 36 00 65 94
italia@gentaur.com | Gentaur