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Key Documents

17-10530

Sigma-Aldrich

ChIPAb+ MED1 Antibody - ChIP Validated Antibody and Primer Set

from rabbit, purified by affinity chromatography

Synonim(y):

CRSP1 ChIP, CRSP1, TRAP220, MED1, TRIP-2, ARC205, Mediator of RNA polymerase II transcription subunit 1, Activator-recruited cofactor 205 kDa component, Mediator complex subunit 1, p53 regulatory protein RB18A, Peroxisome proliferator-activated receptor-binding prot, Chip Antibody and primer set, MED1 ChIP, TRIP-2 ChIP

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About This Item

Kod UNSPSC:
12352203
eCl@ss:
32160702
NACRES:
NA.32

pochodzenie biologiczne

rabbit

Poziom jakości

forma przeciwciała

affinity purified immunoglobulin

klon

polyclonal

oczyszczone przez

affinity chromatography

reaktywność gatunkowa

human, mouse

producent / nazwa handlowa

ChIPAb+
Upstate®

metody

ChIP: suitable
western blot: suitable

numer dostępu NCBI

numer dostępu UniProt

Warunki transportu

dry ice

Opis ogólny

All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ MED1 (CRSP1/TRAP220) set includes a MED1 antibody, a Normal Rabbit IgG, and positive control primers which amplify 85 bp of a human prostate-specific antigen (PSA) enhancer region. The MED1 antibody and negative control IgG are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of MED1 associated chromatin.

Specyficzność

Wide species reactivity expected based upon conservation of immunogen sequence.

Immunogen

Epitope: Human MED1 (a.a. 1525-1581).
Recombinant protein corresponding to human MED1 (a.a. 1525-1581).

Zastosowanie

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from hormone starved (72H), then androgen agonist induced (1 µM CI-4AS-1 overnight) LNCaP cells (~1e6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Rabbit IgG (Part No. PP64B), or 2 µg Anti-MED1 (Part No.CS207349) and the Magna ChIP HiSens (Cat. # 17-10460). Successful immunoprecipitation of MED1 associated DNA fragments was verified by qPCR using ChIP Primers, PSA (Part No. CS207340) as a positive locus, and ChIP Primers, β-globin (Part No. CS200591) as a negative locus.
Please refer to the Magna ChIP HiSens (Cat. # 17-10460) or EZ-MagnaChIP HiSens(Cat. # 17-10461) protocol for experimental details.

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from hormone starved (72H), then androgen agonist induced (1 µM CI-4AS-1 overnight) LNCaP cells (1e5 and 1e6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Rabbit IgG (Part No. PP64B), or 2 µg Anti-MED1 (Part No. CS207349) and the Magna ChIP HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of MED1 associated DNA fragments was verified by qPCR using ChIP Primers, PSA (Part No. CS207340).
Please refer to the MagnaChIP HiSens (Cat. # 17-10460) or EZ-MagnaChIP HiSens (Cat. # 17-10461) protocol for experimental details.

Western Blotting Analysis:
0.04 µg/mL of this antibody detected MED1 on 5, 15 and 50 µg of HeLa whole cell lysate.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Transcription Factors
Use ChIPAb+ MED1 ChIP Validated rabbit polyclonal antibody for the detection of MED also known as Mediator of RNA polymerase II transcription subunit 1 & CRSP1/TRAP220.

Opakowanie

25 assays per set. Recommended use: 2.0 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).

Jakość

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from hormone starved (72h), then androgen agonist induced (1 µM CI-4AS-1 overnight) LNCaP cells (~1e6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Rabbit IgG (Part No. PP64B), or 2 µg Anti-MED1 (Part No.CS207349) and the Magna ChIP® HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of MED1 associated DNA fragments was verified by qPCR using ChIP Primers, PSA (Part No. CS207340).
Please refer to the MagnaChIP HiSens (Cat. # 17-10460) or EZ-MagnaChIP HiSens (Cat. # 17-10461) protocol for experimental details.

Opis wartości docelowych

~168 kDa based on sequence. Higher mw up to 220 kDa observed on western blots.

Postać fizyczna

Anti-MED1 (Rabbit Polyclonal), Part No. CS207349. One vial containing 50 µg affinity purified rabbit polyclonal in 72 uL buffer containing Tris-citrate/phosphate buffer (pH 7 to 8) and 0.09% sodium azide, before the addition of glycerol to 30% (0.7 mg/mL final). Store at -20°C.

Normal Rabbit IgG, Part No. PP64B. One vial containing 125 μg of purified Rabbit IgG in 125 μL storage buffer containing 0.05% sodium azide. Store at -20°C.

ChIP Primers, PSA. Part No. CS207340. One vial containing 75 μL of 5 μM of each primer specific for human prostate-specific antigen enhancer region (chr19:51354179-51354263, hg19 build). Store at -20°C.
FOR: GCC TGG ATC TGA GAG AGA TAT CAT C
REV: ACA CCT TTT TTT TTC TGG ATT GTT G

Przechowywanie i stabilność

Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Komentarz do analizy

Control
Controls
Includes normal rabbit IgG and primers specific for a human prostate-specific antigen enhancer region.

Inne uwagi

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Informacje prawne

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Oświadczenie o zrzeczeniu się odpowiedzialności

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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Kod klasy składowania

10 - Combustible liquids


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