CNOT9 Polyclonal antibody

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  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.

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  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.

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  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.

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  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.

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  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).

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  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).

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  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).

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  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).

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  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).

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

Cat No. 22503-1-AP

Publications(19)

Reviews (2)

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

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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

• 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 17 publications below
• IF: See 2 publications below
• IP: See 1 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: CNOT9 fusion protein Ag18197
• 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: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: Q92600
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

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.

Publications

Species	Application	Title
human	WB	Cell; Phase Transitions in the Assembly and Function of Human miRISC.; Authors - Jessica Sheu-Gruttadauria
human	WB	Mol Cell; Mechanism of ribosome-associated mRNA degradation during tubulin autoregulation; Authors - Markus Höpfler ; KD Validated
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
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
human	WB,IP	Nat Commun; RNF219 attenuates global mRNA decay through inhibition of CCR4-NOT complex-mediated deadenylation.; Authors - Fabian Poetz ; KD Validated
human	WB	Nat Commun; TASOR epigenetic repressor cooperates with a CNOT1 RNA degradation pathway to repress HIV.; Authors - Roy Matkovic

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Roy (Verified Customer) (06-13-2024)

Works well at 1/1000 at 4°C overnight to detect CNOT9 by WB. (also validated through siRNA-mediated KD of CNOT9)

• Applications: Western Blot
• Primary Antibody Dilution: 1/1000
• Cell Tissue Type: HeLa

FH

Abe (Verified Customer) (04-23-2024)

Worked well at 1:100 overnight at 4c.

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:100
• Cell Tissue Type: RPE-1