CNOT1 Polyclonal antibody

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  WB analysis of HeLa using 14276-1-AP

  WB result of CNOT1 antibody (14276-1-AP; 1:300; incubated at room temperature for 1.5 hours) with sh-Control and sh-CNOT1 transfected HeLa cells.

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  WB analysis of human brain using 14276-1-AP

  human brain tissue were subjected to SDS PAGE followed by western blot with 14276-1-AP (CNOT1 antibody) at dilution of 1:800 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse kidney using 14276-1-AP

  mouse kidney tissue were subjected to SDS PAGE followed by western blot with 14276-1-AP (CNOT1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse spleen using 14276-1-AP

  mouse spleen tissue were subjected to SDS PAGE followed by western blot with 14276-1-AP (CNOT1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK293 cells using 14276-1-AP

  WB result from Dr. Vishram Kedar. Lysates from transfected HEK293 cells as indicated in the figure above, were subjected to the western bloting using rabbit CNOT1 antibody (Cat # 14276-1-AP) from Protein-Tech group, Inc, at dilution 1:1000 in 5% non-fat dry milk/TBST, followed by goat anti-rabbit HRP (Bio-RAD) at the dilution of 1:10,000 in 5% non-fat dry milk/TBST. Only bands of the size of 174 kDa were seen. 267 kDa bands of CNOT1 were not visible in this short exposure.

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  WB analysis of mouse thymus using 14276-1-AP

  mouse thymus tissue were subjected to SDS PAGE followed by western blot with 14276-1-AP (CNOT1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse kidney using 14276-1-AP

  mouse kidney tissue were subjected to SDS PAGE followed by western blot with 14276-1-AP (CNOT1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 14276-1-AP

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 14276-1-AP (CNOT1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse brain using 14276-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 14276-1-AP (CNOT1 antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse heart using 14276-1-AP

  mouse heart tissue were subjected to SDS PAGE followed by western blot with 14276-1-AP (CNOT1 antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.

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  IP experiment of mouse kidney using 14276-1-AP

  IP result of anti-CNOT1 (IP:14276-1-AP, 5ug; Detection:14276-1-AP 1:500) with mouse kidney tissue lysate 4000ug.

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  IP experiment of HeLa using 14276-1-AP

  IP result of anti-CNOT1 (IP:14276-1-AP, 4ug; Detection:14276-1-AP 1:600) with HeLa cells lysate 1085 ug.

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  IHC staining of human heart using 14276-1-AP

  Immunohistochemical analysis of paraffin-embedded human heart tissue slide using 14276-1-AP (CNOT1 Antibody) at dilution of 1:200 (under 10x lens).

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  IHC staining of human spleen using 14276-1-AP

  Immunohistochemical analysis of paraffin-embedded human spleen using 14276-1-AP (CNOT1 antibody) at dilution of 1:50 (under 10x lens).

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  IHC staining of human spleen using 14276-1-AP

  Immunohistochemical analysis of paraffin-embedded human spleen using 14276-1-AP (CNOT1 antibody) at dilution of 1:50 (under 40x lens).

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  IHC staining of human kidney using 14276-1-AP

  Immunohistochemical analysis of paraffin-embedded human kidney using 14276-1-AP (CNOT1 antibody) at dilution of 1:50 (under 10x lens).

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  IHC staining of human kidney using 14276-1-AP

  Immunohistochemical analysis of paraffin-embedded human kidney using 14276-1-AP (CNOT1 antibody) at dilution of 1:50 (under 40x lens).

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  IHC staining of human heart using 14276-1-AP

  Immunohistochemical analysis of paraffin-embedded human heart slide using 14276-1-AP (CNOT1 Antibody) at dilution of 1:50.

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  IF Staining of HEK-293 using 14276-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed HEK-293 cells using CNOT1 antibody (14276-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L).

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  IF Staining of HEK-293 using 14276-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed HEK-293 cells using CNOT1 antibody (14276-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L).

• Featured Product
• KD/KO Validated

CNOT1 Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Cat No. 14276-1-AP

Publications(84)

Reviews (6)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (1)

Applications

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

Synonyms

hNOT1, CDC39, CCR4-NOT transcription complex subunit 1, CCR4-associated factor 1, AD-005

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Tested Applications

• Positive WB detected in: human brain tissue, HEK293 cells, mouse kidney tissue, mouse spleen tissue, mouse thymus tissue, HEK-293 cells, mouse brain tissue, mouse heart tissue, HeLa cells
• Positive IP detected in: mouse kidney tissue, HeLa cells
• Positive IHC detected in: human heart tissue, human spleen tissue, human kidney 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: HEK-293 cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:1000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500

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 16 publications below
• WB: See 76 publications below
• IHC: See 2 publications below
• IF: See 6 publications below
• IP: See 5 publications below
• CoIP: See 3 publications below
• ChIP: See 1 publications below
• RIP: See 1 publications below

Product Information

14276-1-AP targets CNOT1 in WB, IHC, IF/ICC, IP, CoIP, chIP, RIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, pig
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: CNOT1 fusion protein Ag5623
• Full Name: CCR4-NOT transcription complex, subunit 1
• Calculated Molecular Weight: 267 kDa
• Observed Molecular Weight: 240-250 kDa, 174 kDa
• GenBank Accession Number: BC040523
• Gene Symbol: CNOT1
• Gene ID (NCBI): 23019
• RRID: AB_10888627
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: A5YKK6
• 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

CNOT1 is a component of CCR4-NOT protein complex, which is a regulator of RNA polymerase II transcription, acts as a transcription repressor. CCR4-NOT complex could participate in transcription as well as mRNA degradation. It's highly expressed in brain, heart, thymus, but weak in skeletal muscle and colon. CNOT1 undergoes alternative splicing to produce four isoforms.This is a rabbit polyclonal antibody raised against part chain of C-terminal CNOT1 of human origin. CNOT1 exsits as many isoforms and molecular weight of isoforms are 267, 241 and 173 kDa.

Publications

Species	Application	Title
human	WB	Nature; DND1 maintains germline stem cells via recruitment of the CCR4-NOT complex to target mRNAs.; Authors - Masashi Yamaji
human	WB	Cell; Roquin promotes constitutive mRNA decay via a conserved class of stem-loop recognition motifs.; Authors - Leppek Kathrin K
mouse	WB	Cell; The Mammalian Ribo-interactome Reveals Ribosome Functional Diversity and Heterogeneity.; Authors - Deniz Simsek
human	WB	Nat Cancer; SULT1A1-dependent sulfonation of alkylators is a lineage-dependent vulnerability of liver cancers; Authors - Lei Shi
	WB	Mol Cell; An Epstein-Barr virus protein interaction map reveals NLRP3 inflammasome evasion via MAVS UFMylation; Authors - Stephanie Pei Tung Yiu
human	WB	Mol Cell; Mechanism of ribosome-associated mRNA degradation during tubulin autoregulation; Authors - Markus Höpfler ; KD Validated

Reviews

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

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FH

Aditya (Verified Customer) (06-20-2024)

Exactly how I would expect an antibodty to behave. Excellent. There is a nonspecific band at 100KDa, but this is probably some variant of CNOT1, so cant fault this at all.

• Applications: Western Blot
• Primary Antibody Dilution: 1:500
• Cell Tissue Type: hek293

FH

Roy (Verified Customer) (06-12-2024)

It does detect CNOT1 by WB from HeLa extracts ( 1/1000 dilution - Overnight incubation at 4°C). It works very well to Immunoprecipitate CNOT1 from both cytoplasmic and nuclear fractions of HeLa cells (4µg /IP)

• Applications: Western Blot, Immunoprecipitation
• Primary Antibody Dilution: 1/1000 (WB)
• Cell Tissue Type: HeLa

FH

aditya (Verified Customer) (10-25-2022)

This is brilliant. We use this for cnot1 pull downs and then use a seperate cnot1 antibody to image on the western. Great for identifying interacting partners.

• Applications: Western Blot, Protein purification
• Primary Antibody Dilution: 1:500
• Cell Tissue Type: Hek293

FH

Tobias (Verified Customer) (10-25-2022)

• Applications: Western Blot, Immunoprecipitation

FH

Parisa (Verified Customer) (10-17-2022)

good quality, high sensitivity

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Hek293

FH

Will (Verified Customer) (01-31-2019)

good antibody, would use again

• Applications: Western Blot,