CNOT4 Recombinant antibody

CNOT4 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

240559E1

Cat no : 83615-4-RR

Synonyms

E3 ubiquitin-protein ligase CNOT4, CLONE243, CCR4-NOT transcription complex subunit 4, CCR4-associated factor 4, CCR4 associated factor 4

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Validation Data Gallery

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  WB analysis using 83615-4-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 83615-4-RR (CNOT4 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IHC staining of human ovary cancer using 83615-4-RR

  Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue slide using 83615-4-RR (CNOT4 antibody) at dilution of 1:300 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 83615-4-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using CNOT4 antibody (83615-4-RR, Clone: 240559E1 ) at dilution of 1:250 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  FC experiment of HeLa using 83615-4-RR

  1x10^6 HeLa cells were intracellularly stained with 0.25 ug Cnot4 Recombinant Antibody (83615-4-RR, Clone:240559E1) and APC-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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  Affinity and Kinetic Characterization of 83615-4-RR

  Biolayer interferometry (BLl) kinetic assays of 83615-4-RR against Human CNOT4 were performed. The affinity constant is 0.485 nM.

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Tested Applications

• Positive WB detected in: MCF-7 cells, HEK-293T cells, HeLa cells, K-562 cells, Jurkat cells, NIH/3T3 cells
• Positive IHC detected in: human ovary cancer 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: HeLa cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:150-1:600
• Immunofluorescence (IF)/ICC: IF/ICC : 1:125-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

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.

Product Information

The immunogen of 83615-4-RR is CNOT4 Fusion Protein expressed in E. coli.

• Tested Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: CNOT4 fusion protein Ag3163
• Full Name: CCR4-NOT transcription complex, subunit 4
• Calculated Molecular Weight: 575 aa, 64 kDa
• Observed Molecular Weight: 64-85 kDa
• GenBank Accession Number: BC035590
• Gene Symbol: CNOT4
• Gene ID (NCBI): 4850
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• 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

CNOT4, also known as CCR4-NOT transcription complex subunit 4 , has E3 ubiquitin ligase activity, and promotes ubiquitination and degradation of target proteins. CNOT4 is involved in activation of the JAK/STAT pathway. CNOT4 exist many isoforms and the range of its molecular weight from 48 to 84 kDa.