RIPAb+ SUZ12 - RIP Validated Antibody and Primer Set

SUZ12 is a polycomb group (PcG) protein. PcG proteins act by forming multiprotein complexes, which are required to maintain the transcriptionally repressive state of homeotic genes throughout development. PcG proteins are not required to initiate repression, but to maintain it during later stages of development. They probably act via the methylation of histones, rendering chromatin heritably changed in its expressibility. Component of the PRC2 complex, which methylates ′Lys-9′ and ′Lys-27′ residues of histone H3.

SUZ12

Full length, recombinant human SUZ12.

Immunoprecipitation from RIP lysate:
Representative lot data.
RIP lysate from HeLa cells (~2 X 10E7 cell equivalents per IP) was subjected to immunoprecipitation using either 5 µg of a normal mouse IgG (Cat. #CS200621), or 5 µg of Anti-SUZ12 antibody (Cat. # CS203201).
20 percent of the precipitated proteins (lane 1: mouse IgG, lane 2: SUZ12) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-SUZ12 antibody (Cat. # CS203201, 1:1000). Proteins were visualized using One-Step IP-Western kit (GenScript Cat. # L00232)
Arrow indicates SUZ12 (Figure 2).

Western Blot Analysis:
Representative lot data.
Lysates from HeLa cells were resolved by electrophoresis, transferred to PVDF and probed with anti-SUZ12 (1:1,000 dilution).
Proteins were visualized using goat anti-mouse secondary antibody conjugated to HRP and chemiluminescence detection.
Arrow SUZ12 (~95 kDa) (Figure 3).

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Chromatin Biology

This RIPAb+ SUZ12 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).

RNA Binding Protein Immunoprecipitation:
RIP Lysate prepared from HeLa cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using either 5 µg of a normal mouse IgG , or 5 µg of Anti-SUZ12 antibody and the Magna RIP^® RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of SUZ12-associated RNA was verified by qPCR using RIP Primers lincSFPQ, (Figure 1)
Please refer to the Magna RIP (Cat. # 17-700) or EZ-Magna RIP (Cat. # 17-701) protocol for experimental details.

95 kDa

Anti-SUZ12 (Mouse Monoclonal) -
One vial containing 50 μg of protein A purified antibody in PBS containing 0.1% azide, before the addition of 30% glycerol. Store at -20°C.

Normal Mouse IgG -
One vial containing 125 µg of purified mouse IgG in 125 µL of storage buffer containing 0.1% sodium azide. Store at -20°C.

RIP Primers, lincSFPQ -
One vial containing 75 μL of 5 μM of each primer specific for the human lincRNA SFPQ. Store at -20°C.
FOR: TCG TAC TGT TAG GCC CTT GG
REV: AAC CTT GCA TGA AGA GCA CC

Format: Purified

Protein A purified

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, 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. Note: Variabillity in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Control
Includes negative control normal mouse IgG antibody and control primers specific for the cDNA of human lincRNA SFPQ.

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

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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.