Validation Data Gallery

Filter: AllWBIPIHCIF/ICC

WB analysis of MCF-7 using 22503-1-AP
WB result of CNOT9 antibody (22503-1-AP; 1:2000; incubated at room temperature for 1.5 hours) with sh-Control and sh-CNOT9 transfected MCF-7 cells.
WB analysis using 22503-1-AP
Various lysates were subjected to SDS PAGE followed by western blot with 22503-1-AP (CNOT9 antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.
WB analysis of mouse thymus using 22503-1-AP
mouse thymus tissue were subjected to SDS PAGE followed by western blot with 22503-1-AP (RQCD1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
IP experiment of MCF-7 using 22503-1-AP
IP result of anti-CNOT9 (IP:22503-1-AP, 4ug; Detection:22503-1-AP 1:1000) with MCF-7 cells lysate 1280ug.
IHC staining of human stomach cancer using 22503-1-AP
Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 22503-1-AP (CNOT9 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human stomach cancer using 22503-1-AP
Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 22503-1-AP (CNOT9 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of rat colon using 22503-1-AP
Immunohistochemical analysis of paraffin-embedded rat colon tissue slide using 22503-1-AP (CNOT9 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of rat colon using 22503-1-AP
Immunohistochemical analysis of paraffin-embedded rat colon tissue slide using 22503-1-AP (CNOT9 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IF Staining of HepG2 using 22503-1-AP
Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using CNOT9 antibody (22503-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).
IF Staining of hTERT-RPE1 using 22503-1-AP
Immunofluorescent analysis of (4% PFA) fixed hTERT-RPE1 cells using CNOT9 antibody (22503-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

Featured Product
KD/KO Validated

CNOT9 Polyclonal antibody

CNOT9 Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Cat No. 22503-1-AP

Publications(20)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, IP, CoIP, ELISA

Synonyms

RQCD1, Rcd-1, RCD1, Rcd 1, Cell differentiation protein RQCD1 homolog

Formulation: PBS, Azide, Glycerol

PBS, Azide, Glycerol

Conjugate: Unconjugated

Unconjugated

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (20)
Protocols

データシート［PDF］を印刷

MSDS

Tested Applications

Positive WB detected in	HeLa cells, MCF-7 cells, mouse thymus tissue, HepG2 cells
Positive IP detected in	MCF-7 cells
Positive IHC detected in	human stomach cancer tissue, rat colon tissue **Note: suggested antigen retrieval with TE buffer pH 9.0; () Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0***
Positive IF/ICC detected in	HepG2 cells, hTERT-RPE1 cells

Recommended dilution

Application	Dilution
Western Blot (WB)	WB : 1:1000-1:4000
Immunoprecipitation (IP)	IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunohistochemistry (IHC)	IHC : 1:500-1:2000
Immunofluorescence (IF)/ICC	IF/ICC : 1:200-1:800
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Published Applications

KD/KO	See 2 publications below
WB	See 18 publications below
IF	See 2 publications below
IP	See 2 publications below
CoIP	See 2 publications below

Product Information

22503-1-AP targets CNOT9 in WB, IHC, IF/ICC, IP, CoIP, ELISA applications and shows reactivity with human, mouse, rat samples.

Tested Reactivity	human, mouse, rat
Cited Reactivity	human, mouse
Host / Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	CatNo: Ag18197 Product name: Recombinant human RQCD1 protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 28-269 aa of BC007102 Sequence: MHSLATAAPVPTTLAQVDREKIYQWINELSSPETRENALLELSKKRESVPDLAPMLWHSFGTIAALLQEIVNIYPSINPPTLTAHQSNRVCNALALLQCVASHPETRSAFLAAHIPLFLYPFLHTVSKTRPFEYLRLTSLGVTGALVKTDEQEVINFLLTTEIIPLCLRIMESGSELSKTVATFILQKILLDDTGLAYICQTYERFSHVAMILGKMVLQLSKEPSARLLKHVVRCYLRLSDN 相同性解析による交差性が予測される生物種
Full Name	RCD1 required for cell differentiation1 homolog (S. pombe)
Calculated molecular weight	299 aa, 34 kDa
Observed molecular weight	30-40 kDa
GenBank accession number	BC007102
Gene Symbol	CNOT9
Gene ID (NCBI)	9125
RRID	AB_11232413
Conjugate	Unconjugated
Form
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	Q92600
Storage Buffer	PBS with 0.02% sodium azide and 50% glycerol{{ptg:BufferTemp}}7.3
Storage Conditions	Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

Background Information

RQCD1, also known as CNOT9, is a highly conserved transcriptional cofactor predominantly expressed in actively diffierentiating tissues like testis and thymus. It is a component of the CCR4-NOT complex, the major effector complex in miRNA-mediated gene silencing. Overexpression of RQCD1 has been reported in serveral tumors, indicating that it may be a potential molecular target for cancer treatment.

Protocols

Product Specific Protocols
IF protocol for CNOT9 antibody 22503-1-AP	Download protocol
IHC protocol for CNOT9 antibody 22503-1-AP	Download protocol
IP protocol for CNOT9 antibody 22503-1-AP	Download protocol
WB protocol for CNOT9 antibody 22503-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

Species	Application	Title
human	WB	Cell Phase Transitions in the Assembly and Function of Human miRISC. Authors - Jessica Sheu-Gruttadauria View Article
human	WB	Mol Cell Mechanism of ribosome-associated mRNA degradation during tubulin autoregulation Authors - Markus Höpfler KD Validated View Article
human	WB	Mol Cell Structural and Biochemical Insights to the Role of the CCR4-NOT Complex and DDX6 ATPase in MicroRNA Repression. Authors - Hansruedi Mathys View Article
human	WB	Mol Cell A DDX6-CNOT1 Complex and W-Binding Pockets in CNOT9 Reveal Direct Links between miRNA Target Recognition and Silencing. Authors - Ying Chen View Article
human	WB,IP	Nat Commun RNF219 attenuates global mRNA decay through inhibition of CCR4-NOT complex-mediated deadenylation. Authors - Fabian Poetz KD Validated View Article
human	WB	Nat Commun TASOR epigenetic repressor cooperates with a CNOT1 RNA degradation pathway to repress HIV. Authors - Roy Matkovic View Article

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis of MCF-7 using 22503-1-AP

WB analysis using 22503-1-AP

WB analysis of mouse thymus using 22503-1-AP

IP experiment of MCF-7 using 22503-1-AP

IHC staining of human stomach cancer using 22503-1-AP

IHC staining of human stomach cancer using 22503-1-AP

IHC staining of rat colon using 22503-1-AP

IHC staining of rat colon using 22503-1-AP

IF Staining of HepG2 using 22503-1-AP

IF Staining of hTERT-RPE1 using 22503-1-AP

CNOT9 Polyclonal antibody

CNOT9 Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

-/ -

Tested Applications

Recommended dilution

Published Applications

Product Information

Background Information

Protocols

Publications

Phase Transitions in the Assembly and Function of Human miRISC.

Mechanism of ribosome-associated mRNA degradation during tubulin autoregulation

Structural and Biochemical Insights to the Role of the CCR4-NOT Complex and DDX6 ATPase in MicroRNA Repression.

A DDX6-CNOT1 Complex and W-Binding Pockets in CNOT9 Reveal Direct Links between miRNA Target Recognition and Silencing.

RNF219 attenuates global mRNA decay through inhibition of CCR4-NOT complex-mediated deadenylation.

TASOR epigenetic repressor cooperates with a CNOT1 RNA degradation pathway to repress HIV.