Invitrogen

MNAT1 Polyclonal Antibody

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Datasheet

Protocols

Questions & Answers

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MNAT1 Antibody in Immunocytochemistry (ICC/IF) — Immunocytochemistry analysis of MNAT1 using anti-MNAT1 antibody (Product # PA5-79690) . MNAT1 was detected in a section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 5μg/mL rabbit anti-MNAT1 antibody (Product # PA5-79690) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

MNAT1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of MNAT1 on paraffin-embedded human testis tissue. Sample was incubated with MNAT1 polyclonal antibody (Product# PA5-79690).

MNAT1 Antibody in Western Blot (WB) — Western blot analysis of MNAT1 in, Lane 1: human Hela whole cell lysates, Lane 2: human A431 whole cell lysates, Lane 3: human A375 whole cell lysates, Lane 4: human MCF-7 whole cell lysates, Lane 5: rat testis tissue lysates, Lane 6: rat L6 whole cell lysates, Lane 7: mouse testis tissue lysates, Lane 8: mouse NIH/3T3 whole cell lysates. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. The membrane was blocked with 5% non-fat milk/TBS for 1. 5 hour at RT. The membrane was incubated with MNAT1 Polyclonal Antibody (Product # PA5-79690) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0. 1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5,000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for MNAT1 at approximately 36 kDa. The expected band size for MNAT1 is at 36 kDa.

MNAT1 Antibody in Flow Cytometry (Flow) — Flow Cytometry of MNAT1 in A431 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with MNAT1 Polyclonal Antibody (Product # PA5-79690) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.

Product Details

PA5-79690

Applications

Tested Dilution

Publications

Western Blot (WB)

0.1-0.5 µg/mL

Immunohistochemistry (Paraffin) (IHC (P))

0.5-1 µg/mL

Immunocytochemistry (ICC/IF)

5 µg/mL

Flow Cytometry (Flow)

1-3 µg/1x10^6 cells

Product Specifications

Species Reactivity

Human, Mouse, Rat

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

E.coli-derived human MNAT1 recombinant protein (Position: L92-A278).

View immunogen

Conjugate

Unconjugated

Form

Lyophilized

Concentration

500 µg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS with 5mg BSA

Contains

0.05mg sodium azide

Storage conditions

-20°C

RRID

AB_2746805

Product Specific Information

Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL.

Target Information

Progression through the cell cycle requires activation of a series of enzymes designated cyclin dependent kinases (Cdks). The monomeric catalytic subunit, Cdk2, a critical enzyme for initiation of cell cycle progression, is completely inactive. Partial activation is achieved by the binding of regulatory cyclins such as cyclin D1, while full activation requires phosphorylation at Thr 160. The enzyme responsible for phosphorylation of Thr 160 in Cdk2 and also Thr 161 in Cdc2 p34, designated Cdk-activating kinase (CAK), has been partially purified and shown to be comprised of a catalytic subunit, a regulatory subunit and a subunit of unknown function. The regulatory subunit is a novel cyclin (cyclin H) and is required for activation of Cdk7. This previously undescribed protein, now termed Mat1, has been cloned as a protein that associates with the cyclin H-Cdk7 nuclear complex at all stages of the cell cycle. Cyclin H-Cdk7-Mat1 complexes display kinase activity towards Cdk activation domains, and the carboxy terminus of RNA polymerase II. Mat1 appears to constitute the first example of an assembly factor, essential for the formation of an active Cdk-cyclin complex.

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

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Bioinformatics

Protein Aliases: AdoMet synthetase; AMS1; CDK-activating kinase assembly factor MAT1; CDK7/cyclin H assembly factor; CDK7/cyclin-H assembly factor; cyclin G1 interacting protein; Cyclin-G1-interacting protein; EC 2.5.1.6; MATA1; Menage a trois; menage a trois 1; menage a trois 1 (CAK assembly factor); menage a trois homolog 1, cyclin H assembly factor; menage a trois-like protein 1 cyclin H assembly factor; MNAT CDK-activating kinase assembly factor 1; p35; p36; RING finger protein 66; RING finger protein MAT1

Gene Aliases: CAP35; E130115E11Rik; MAT1; MNAT1; P36; RNF66; TFB3

UniProt ID: (Human) P51948, (Mouse) P51949

Entrez Gene ID: (Human) 4331, (Mouse) 17420, (Rat) 266713

Molecular Function: chaperone