RIPAb+™ hnRNP C1/C2 - RIP Validated Antibody and Primer Set
Overview
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.
The hnRNP C proteins (C1/C2) are nuclear pre-mRNA-binding proteins that belong to the large RNP motif family of RNA-binding proteins. The hnRNPs are RNA binding proteins that complex with heterogeneous nuclear RNA (hnRNA). These proteins associate with pre-mRNAs in the nucleus and influence pre-mRNA processing and other aspects of mRNA metabolism and transport.
Background Information
The hnRNP C proteins (C1/C2) are nuclear pre-mRNA-binding proteins that belong to the large RNP motif family of RNA-binding proteins. The proteins encoded by this gene act as a tetramer and are involved in the assembly of 40S hnRNP particles. The hnRNPs are RNA binding proteins that complex with heterogeneous nuclear RNA (hnRNA). These proteins aassociate with pre-mRNAs in the nucleus and influence pre-mRNA processing and other aspects of mRNA metabolism and transport. In vivo, hnRNP C proteins have been found to interact with Grb2.
Product Information
Format
Purified
Control
Includes negative control normal mouse IgG antibody and control primers specific for the cDNA of human c-myc 3' UTR.
Presentation
Anti-hnRNP C1/C2 (Mouse Monoclonal), . One vial containing 50 µg of protein G purified mouse IgG1 in 0.1M Tris-glycine, pH 7.4, 0.15M NaCl, 0.05% sodium azide and 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, c-myc 3' UTR, . One vial containing 75 μL of 5 μM of each primer specific for human c-myc 3' UTR. Store at -20°C.
FOR: CAC AAA CTT GAA CAG CTA CGG
REV: GGT GAT TGC TCA GGA CAT TTC
Applications
Application
This RIPAb+ hnRNP C1/C2 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
Western Blotting
RNA Binding Protein Immunoprecipitation (RIP)
Application Notes
Immunoprecipitation from RIP lysate:
Representative lot data.
RIP lysate from HeLa cells (~2 X 10E7 cell equivalents per IP) was subjected to immunoprecipitation using 5 µg of either a normal mouse IgG, (Cat. # CS200621), or 5 µg of Anti-hnRNP C1/C2 antibody (Cat. # CS207305). ten percent of the precipitated proteins (lane 1: normal mouse IgG, lane 2: hnRNP C1/C2) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-hnRNP C1/C2 (Cat. # CS207305, 1:1000). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00231).
Arrow indicates hnRNP C1/C2. (Figure 2).
Automated Microfluidics-based Electrophoretic RNA Separation and Analysis (MFE):
Representative lot data.
RIP Lysate prepared from HeLa cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either 1. normal mouse IgG (Cat. # CS200621), or 2. Anti-hnRNP C1/C2 antibody (Cat. # CS207305) and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of hnRNP C1/C2-associated RNA was verified by automated microfluidics-based electrophoretic RNA separation and analysis. Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details. Electropherograms were generated by plotting fluorescence intensities versus migration times (Figure 3A). The virtual gel view was created from this data (Figure 3B).
Western Blot Analysis:
Representative lot data.
HeLa nuclear extract lysate was probed with Anti-hnRNP C1/C2, clone 4F4 (0.01 µg/mL). Proteins were visualized using a Goat Anti-Mouse IgG secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates hnRNP C1/C2 (~37/39 kDa). (Figure 4).
Note: The calculated molecular weight for C1 is 32.3 kDa and 33.8 kDa for C2.
Biological Information
Immunogen
RNP’s eluted from oligo (dT) cellulose column of human HnRNP C1/C2.
Epitope
Unknown
Concentration
Please refer to the Certificate of Analysis for the lot-specific concentration.
Host
Mouse
Specificity
This antibody recognizes HnRNP C1/C2.
Isotype
IgG2bκ
Species Reactivity
Mouse Human
Species Reactivity Note
Demonstrated to react with human. Predicted to react with mouse based on 100% sequence homology.
This gene belongs to the subfamily of ubiquitously expressed heterogeneous nuclear ribonucleoproteins (hnRNPs). The hnRNPs are RNA binding proteins and they complex with heterogeneous nuclear RNA (hnRNA). These proteins are associated with pre-mRNAs in the nucleus and appear to influence pre-mRNA processing and other aspects of mRNA metabolism and transport. While all of the hnRNPs are present in the nucleus, some seem to shuttle between the nucleus and the cytoplasm. The hnRNP proteins have distinct nucleic acid binding properties. The protein encoded by this gene can act as a tetramer and is involved in the assembly of 40S hnRNP particles. Multiple transcript variants encoding at least two different isoforms have been described for this gene. [provided by RefSeq].
"FUNCTION: Binds pre-mRNA and nucleates the assembly of 40S hnRNP particles. Single HNRNPC tetramers bind 230-240 nucleotides. Trimers of HNRNPC tetramers bind 700 nucleotides. May play a role in the early steps of spliceosome assembly and pre-mRNA splicing. Interacts with poly-U tracts in the 3'-UTR or 5'-UTR of mRNA and modulates the stability and the level of translation of bound mRNA molecules.
SUBUNIT STRUCTURE: Tetramer composed of 3 copies of isoform C1 and 1 copy of isoform C2. Assembly of 3 tetramers with bound pre-mRNA gives rise to a 19S complex that interacts with HNRNPA2B1 tetramers. Component of the 40S hnRNP particle. Identified in the spliceosome C complex, at least composed of AQR, ASCC3L1, C19orf29, CDC40, CDC5L, CRNKL1, DDX23, DDX41, DDX48, DDX5, DGCR14, DHX35, DHX38, DHX8, EFTUD2, FRG1, GPATC1, HNRNPA1, HNRNPA2B1, HNRPA3, HNRNPC, HNRPF, HNRPH1, HNRPK, HNRPM, HNRNPR, HNRNPU, KIAA1160, KIAA1604, LSM2, LSM3, MAGOH, MORG1, PABPC1, PLRG1, PNN, PPIE, PPIL1, PPIL3, PPWD1, PRPF19, PRPF4B, PRPF6, PRPF8, RALY, RBM22, RBM8A, RBMX, SART1, SF3A1, SF3A2, SF3A3, SF3B1, SF3B2, SF3B3, SFRS1, SKIV2L2, SNRPA1, SNRPB, SNRPB2, SNRPD1, SNRPD2, SNRPD3, SNRPE, SNRPF, SNRPG, SNW1, SRRM1, SRRM2, SYF2, SYNCRIP, TFIP11, THOC4, U2AF1, WDR57, XAB2 and ZCCHC8. Interacts with IGF2BP1.
SUBCELLULAR LOCATION: Nucleus. Note: Component of ribonucleosomes.
PTM: Phosphorylated on Ser-260 and Ser-299 in resting cells. Phosphorylated on Ser-253 and on 1 serine residue in the poly-Ser stretch at position 238 in response to hydrogen peroxide.
Sumoylated. Sumoylation reduces affinity for mRNA.
SEQUENCE SIMILARITIES: Belongs to the RRM HNRPC family. RALY subfamily.
Contains 1 RRM (RNA recognition motif) domain.
Product Usage Statements
Quality Assurance
RNA Binding Protein Immunoprecipitation:
Representative lot data.
RIP Lysate prepared from HeLa cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG , or 5 µg of Anti-hnRNP C1/C2 antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of hnRNP C1/C2-associated RNA was verified by qPCR using RIP Primers c-myc 3' UTR, (Figure 1).
Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.
Usage Statement
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.
Storage and Shipping Information
Storage Conditions
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.
Packaging Information
Material Size
10 assays
Material Package
10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).
原厂资料：
Key Spec Table
Species Reactivity
Key Applications
M, H
WB, RIP
Description
Catalogue Number
03-205
Trade Name
Upstate
RIPAb+
Description
RIPAb+™ hnRNP C1/C2 - RIP Validated Antibody and Primer Set
Overview
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.
The hnRNP C proteins (C1/C2) are nuclear pre-mRNA-binding proteins that belong to the large RNP motif family of RNA-binding proteins. The hnRNPs are RNA binding proteins that complex with heterogeneous nuclear RNA (hnRNA). These proteins associate with pre-mRNAs in the nucleus and influence pre-mRNA processing and other aspects of mRNA metabolism and transport.
Background Information
The hnRNP C proteins (C1/C2) are nuclear pre-mRNA-binding proteins that belong to the large RNP motif family of RNA-binding proteins. The proteins encoded by this gene act as a tetramer and are involved in the assembly of 40S hnRNP particles. The hnRNPs are RNA binding proteins that complex with heterogeneous nuclear RNA (hnRNA). These proteins aassociate with pre-mRNAs in the nucleus and influence pre-mRNA processing and other aspects of mRNA metabolism and transport. In vivo, hnRNP C proteins have been found to interact with Grb2.
Product Information
Format
Purified
Control
Includes negative control normal mouse IgG antibody and control primers specific for the cDNA of human c-myc 3' UTR.
Presentation
Anti-hnRNP C1/C2 (Mouse Monoclonal), . One vial containing 50 µg of protein G purified mouse IgG1 in 0.1M Tris-glycine, pH 7.4, 0.15M NaCl, 0.05% sodium azide and 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, c-myc 3' UTR, . One vial containing 75 μL of 5 μM of each primer specific for human c-myc 3' UTR. Store at -20°C.
FOR: CAC AAA CTT GAA CAG CTA CGG
REV: GGT GAT TGC TCA GGA CAT TTC
Applications
Application
This RIPAb+ hnRNP C1/C2 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
Western Blotting
RNA Binding Protein Immunoprecipitation (RIP)
Application Notes
Immunoprecipitation from RIP lysate:
Representative lot data.
RIP lysate from HeLa cells (~2 X 10E7 cell equivalents per IP) was subjected to immunoprecipitation using 5 µg of either a normal mouse IgG, (Cat. # CS200621), or 5 µg of Anti-hnRNP C1/C2 antibody (Cat. # CS207305). ten percent of the precipitated proteins (lane 1: normal mouse IgG, lane 2: hnRNP C1/C2) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-hnRNP C1/C2 (Cat. # CS207305, 1:1000). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00231).
Arrow indicates hnRNP C1/C2. (Figure 2).
Automated Microfluidics-based Electrophoretic RNA Separation and Analysis (MFE):
Representative lot data.
RIP Lysate prepared from HeLa cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either 1. normal mouse IgG (Cat. # CS200621), or 2. Anti-hnRNP C1/C2 antibody (Cat. # CS207305) and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of hnRNP C1/C2-associated RNA was verified by automated microfluidics-based electrophoretic RNA separation and analysis. Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details. Electropherograms were generated by plotting fluorescence intensities versus migration times (Figure 3A). The virtual gel view was created from this data (Figure 3B).
Western Blot Analysis:
Representative lot data.
HeLa nuclear extract lysate was probed with Anti-hnRNP C1/C2, clone 4F4 (0.01 µg/mL). Proteins were visualized using a Goat Anti-Mouse IgG secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates hnRNP C1/C2 (~37/39 kDa). (Figure 4).
Note: The calculated molecular weight for C1 is 32.3 kDa and 33.8 kDa for C2.
Biological Information
Immunogen
RNP’s eluted from oligo (dT) cellulose column of human HnRNP C1/C2.
Epitope
Unknown
Concentration
Please refer to the Certificate of Analysis for the lot-specific concentration.
Host
Mouse
Specificity
This antibody recognizes HnRNP C1/C2.
Isotype
IgG2bκ
Species Reactivity
Mouse Human
Species Reactivity Note
Demonstrated to react with human. Predicted to react with mouse based on 100% sequence homology.
This gene belongs to the subfamily of ubiquitously expressed heterogeneous nuclear ribonucleoproteins (hnRNPs). The hnRNPs are RNA binding proteins and they complex with heterogeneous nuclear RNA (hnRNA). These proteins are associated with pre-mRNAs in the nucleus and appear to influence pre-mRNA processing and other aspects of mRNA metabolism and transport. While all of the hnRNPs are present in the nucleus, some seem to shuttle between the nucleus and the cytoplasm. The hnRNP proteins have distinct nucleic acid binding properties. The protein encoded by this gene can act as a tetramer and is involved in the assembly of 40S hnRNP particles. Multiple transcript variants encoding at least two different isoforms have been described for this gene. [provided by RefSeq].
"FUNCTION: Binds pre-mRNA and nucleates the assembly of 40S hnRNP particles. Single HNRNPC tetramers bind 230-240 nucleotides. Trimers of HNRNPC tetramers bind 700 nucleotides. May play a role in the early steps of spliceosome assembly and pre-mRNA splicing. Interacts with poly-U tracts in the 3'-UTR or 5'-UTR of mRNA and modulates the stability and the level of translation of bound mRNA molecules.
SUBUNIT STRUCTURE: Tetramer composed of 3 copies of isoform C1 and 1 copy of isoform C2. Assembly of 3 tetramers with bound pre-mRNA gives rise to a 19S complex that interacts with HNRNPA2B1 tetramers. Component of the 40S hnRNP particle. Identified in the spliceosome C complex, at least composed of AQR, ASCC3L1, C19orf29, CDC40, CDC5L, CRNKL1, DDX23, DDX41, DDX48, DDX5, DGCR14, DHX35, DHX38, DHX8, EFTUD2, FRG1, GPATC1, HNRNPA1, HNRNPA2B1, HNRPA3, HNRNPC, HNRPF, HNRPH1, HNRPK, HNRPM, HNRNPR, HNRNPU, KIAA1160, KIAA1604, LSM2, LSM3, MAGOH, MORG1, PABPC1, PLRG1, PNN, PPIE, PPIL1, PPIL3, PPWD1, PRPF19, PRPF4B, PRPF6, PRPF8, RALY, RBM22, RBM8A, RBMX, SART1, SF3A1, SF3A2, SF3A3, SF3B1, SF3B2, SF3B3, SFRS1, SKIV2L2, SNRPA1, SNRPB, SNRPB2, SNRPD1, SNRPD2, SNRPD3, SNRPE, SNRPF, SNRPG, SNW1, SRRM1, SRRM2, SYF2, SYNCRIP, TFIP11, THOC4, U2AF1, WDR57, XAB2 and ZCCHC8. Interacts with IGF2BP1.
SUBCELLULAR LOCATION: Nucleus. Note: Component of ribonucleosomes.
PTM: Phosphorylated on Ser-260 and Ser-299 in resting cells. Phosphorylated on Ser-253 and on 1 serine residue in the poly-Ser stretch at position 238 in response to hydrogen peroxide.
Sumoylated. Sumoylation reduces affinity for mRNA.
SEQUENCE SIMILARITIES: Belongs to the RRM HNRPC family. RALY subfamily.
Contains 1 RRM (RNA recognition motif) domain.
Product Usage Statements
Quality Assurance
RNA Binding Protein Immunoprecipitation:
Representative lot data.
RIP Lysate prepared from HeLa cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG , or 5 µg of Anti-hnRNP C1/C2 antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of hnRNP C1/C2-associated RNA was verified by qPCR using RIP Primers c-myc 3' UTR, (Figure 1).
Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.
Usage Statement
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.
Storage and Shipping Information
Storage Conditions
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.
Packaging Information
Material Size
10 assays
Material Package
10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).
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