半岛bd体育手机客户端 说明
一般描述
A+U Rich RNA Binding Factor (AUF1) is comprised of four isoforms of 37, 40, 42, and 45 kDa. Although the role of each isoform has yet to be fully characterized, a direct correlation has been observed between each AUF1 isoform’s binding affinity and its RNA destabilizing activity toward different AREs (A + U rich element in the 3’ untranslated region), with the isoforms p37 and p42 being the most effective.
AUF1 is a bcl2 mRNA binding protein and potentially all its isoforms are able to form complexes with the bcl2 ARE. ARE mediated bcl2 mRNA downregulation during apoptosis involves AUF1 and suggest different roles for its four isoforms.
RIPAb+ antibodies are evaluated using the RNA Binding Protein Immunoprecipitation (RIP) assay. Each RIPAb+ antibody set includes a negative control antibody to ensure specificity of the RIP reaction and is verified for the co-immunoprecipitation of RNA associated specifically with the immunoprecipitated RNA binding protein of interest. Where appropriate, the RIPAb+ set also includes quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully co-precipitating the specific RNA targets, such as messenger RNAs. The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. If a target specific assay is not provided, the RIPAb+ kit is validated using an automated microfluidics-based assay by enrichment of detectable RNA over control immunoprecipitation.
特异性
Predicted cross-reactivity with mouse and rat based on sequence homology.
AUF1 isoforms at 37, 40, 42 and 45 kDa.
免疫原
Human AUF1 purified by Ni2+affinity column.
应用
Immunoprecipitation from RIP lysate:Representative lot data.
RIP lysate from HeLa cells (2 X 106cell equivalents per IP) was subjected to immunoprecipitation using 0.5 ?g of either a normal rabbit IgG or Anti-AUF1 antibody. Precipitated proteins (Lane 1: rabbit IgG, Lane 2: anti-AUF1) and HeLa whole cell lysate (Lane 3) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-AUF1 antibody (1.0 ?g/mL).
Proteins were visualized using One-Step
Western Blot Analysis:0.01-1 ?g/mL of a previous lot detected AUF1 in RIPA lysates from HeLa nuclear extract and A431 cells.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
RNA Metabolism & Binding Proteins
RNA Binding Protein (RBP)
This RIPAb+ AUF1 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
包装
10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).
质量
RNA Binding Protein Immunoprecipitation:
RIP Lysate prepared from HeLa cells (2 X 107cell equivalents per IP) were subjected to immunoprecipitation using 5 ?g of either a normal rabbit IgG or 5 ?g of Anti-AUF1 antibody and the Magna RIP?RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of AUF1-associated RNA was verified by qPCR using RIP Primers FOS (Please see figures).
Please refer to the Magna RIP
目标描述
37, 40, 42 and 45 kDa
外形
Format: Purified
Anti-AUF1 (Rabbit Polyclonal). One vial containing 50 μg of protein A purified rabbit IgG in 50 μL of 70% storage buffer (0.1M Tris-glycine, pH 7.4, 0.15M NaCl, 0.05% sodium azide) and 30% glycerol. Store at -20°C.
Normal Rabbit IgG. One vial containing 125 μg Rabbit IgG in 125 μL storage buffer containing 0.05% sodium azide. Store at -20°C.
RIP Primers, FOS. One vial containing 75 μL of 5 μM of each primer specific for human FOS. Store at -20°C.
FOR: GAG AGC TGG TAG TTA GTA GCA TGT TGA
REV: AAT TCC AAT AAT GAA CCC AAT AGA TTA GTT A
储存及稳定性
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: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
分析说明
Control
Includes negative control rabbit IgG antibody and control primers specific for human FOS.
法律信息
MAGNA RIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
免责声明
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.
基本信息
| eCl@ss | 32160702 |
| NACRES | NA.32 |
半岛bd体育手机客户端 性质
| 生物来源 | rabbit |
| 质量水平 | 100 |
| 抗体形式 | purified immunoglobulin |
| 克隆 | polyclonal |
| species reactivity | rat, human, mouse |
| manufacturer/tradename | RIPAb+ Upstate? |
| technique(s) | RIP: suitable immunoprecipitation (IP): suitable western blot: suitable |
| NCBI登记号 | NP_001003810.1 |
| UniProt登记号 | Q14103 |
| 运输 | dry ice |
| Gene Information | human ... HNRNPD(3184) |
安全信息
| 储存分类代码 | 10 - Combustible liquids |
Sigma-Aldrich
m.cnreagent.com
