Nucleolin/C23 Recombinant antibody

Nucleolin/C23 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

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

Conjugate

Unconjugated

CloneNo.

240029F8

Cat no : 83380-1-RR

Synonyms

C23, nucleolin, Protein C23, NCL, FLJ45706

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Validation Data Gallery

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  WB analysis of HeLa using 83380-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 83380-1-RR (Nucleolin/C23 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 83380-1-RR

  IP result of anti-Nucleolin/C23 (IP:83380-1-RR, 4ug; Detection:83380-1-RR 1:5000) with HeLa cells lysate 1085 ug.

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  IHC staining of human lung cancer using 83380-1-RR

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 83380-1-RR (Nucleolin/C23 antibody) at dilution of 1:400 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 83380-1-RR

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 83380-1-RR (Nucleolin/C23 antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 83380-1-RR

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using Nucleolin/C23 antibody (83380-1-RR, Clone: 240029F8 ) at dilution of 1:250 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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  FC experiment of HEK-293 using 83380-1-RR

  1x10^6 HEK-293 cells were intracellularly stained with 0.25 ug Nucleolin/C23 Recombinant antibody (83380-1-RR, Clone:240029F8) and APC-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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  Affinity and Kinetic Characterization of 83380-1-RR

  Biolayer interferometry (BLl) kinetic assays of 83380-1-RR against Human Nucleolin/C23 were performed. The affinity constant is 2.47 nM.

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, Jurkat cells, K-562 cells, MCF-7 cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human lung 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: HepG2 cells
• Positive FC (Intra) detected in: HEK-293 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:200-1:800
• 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 83380-1-RR is Nucleolin/C23 Fusion Protein expressed in E. coli.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: Nucleolin/C23 fusion protein Ag0859
• Full Name: nucleolin
• Calculated Molecular Weight: 76 kDa
• Observed Molecular Weight: 100-110 kDa
• GenBank Accession Number: BC006494
• Gene Symbol: NCL
• Gene ID (NCBI): 4691
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• 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

Nucleolin, also known as C23, involved in the control of transcription of ribosomal RNA (rRNA) genes by RNA polymerase I, in nucleocytoplasmic transportation of ribosomal components, and in ribosome maturation and assembly. It associated with intranucleolar chromatin and pre-ribosomal particles, and induced chromatin decondensation by binding to histione H1. Also it has a role in the process of transcriptional elongation. Whilst mammalian nucleolin has a predicted molecular mass of approximately 77 kDa, the apparent molecular mass is between 100 and 110 kDa, and has been attributed to the amino acid composition of the N-terminal domain, which is highly phosphorylated. (PMID: 15925566)