Nucleobindin 1 Polyclonal antibody

Nucleobindin 1 Polyclonal Antibody for WB, IHC, IF/ICC, IF-P, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

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

Conjugate

Unconjugated

Publications

1

Cat no : 10228-1-AP

Synonyms

NUCB1, Nucleobindin-1, NUC, CALNUC

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Validation Data Gallery

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  WB analysis of MCF-7 using 10228-1-AP

  MCF7 cells were subjected to SDS PAGE followed by western blot with 10228-1-AP (nucleobindin 1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of A431 using 10228-1-AP

  A431 cells were subjected to SDS PAGE followed by western blot with 10228-1-AP (nucleobindin 1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of HeLa using 10228-1-AP

  HeLa cells were subjected to SDS PAGE followed by western blot with 10228-1-AP (nucleobindin 1 antibody at dilution of 1:600 incubated at room temperature for 1.5 hours.

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  WB analysis of HepG2 using 10228-1-AP

  HepG2 cells were subjected to SDS PAGE followed by western blot with 10228-1-AP (nucleobindin 1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IP experiment of HepG2 using 10228-1-AP

  IP result of anti-nucleobindin 1 (IP:10228-1-AP, 4ug; Detection:10228-1-AP 1:1000) with HepG2 cells lysate 2400ug.

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  IHC staining of human brain using 10228-1-AP

  Immunohistochemical analysis of paraffin-embedded human brain tissue slide using 10228-1-AP (nucleobindin 1 antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human brain using 10228-1-AP

  Immunohistochemical analysis of paraffin-embedded human brain tissue slide using 10228-1-AP (nucleobindin 1 antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of mouse brain using 10228-1-AP

  Immunofluorescent analysis of (4% PFA) fixed paraffin-embedded mouse brain tissue using nucleobindin 1 antibody (10228-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 10228-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using 10228-1-AP (nucleobindin 1 antibody) at dilution of 1:50 and CoraLite488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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  FC experiment of HepG2 using 10228-1-AP

  1x10^6 HepG2 cells were intracellularly stained with 0.25 ug nucleobindin 1 Polyclonal antibody (10228-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: MCF7 cells, A431 cells, HeLa cells, HepG2 cells
• Positive IP detected in: HepG2 cells
• Positive IHC detected in: human brain 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-P detected in: mouse brain tissue
• Positive IF/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: HepG2 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)-P: IF-P : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• 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.

Published Applications

• WB: See 1 publications below

Product Information

10228-1-AP targets Nucleobindin 1 in WB, IHC, IF/ICC, IF-P, FC (Intra), IP, ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Cited Reactivity: mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: Nucleobindin 1 fusion protein Ag0290
• Full Name: nucleobindin 1
• Calculated Molecular Weight: 55 kDa
• Observed Molecular Weight: 63 kDa
• GenBank Accession Number: BC002356
• Gene Symbol: NUCB1
• Gene ID (NCBI): 4924
• RRID: AB_2153655
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity 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

Nucleobindin-1 (NUCB1), also known as Calnuc, is a highly conserved, multifunctional protein widely expressed in tissues and cells.It has been proposed to contain both calcium- and DNA-binding motifs, including two EF-hands, a basic amino-acid-rich region and a leucine zipper.Western blotting revealed a band at approximately 63 kDa, corresponding to the expected size of the NUCB1 protein(PMID: 15255074)

Publications

Species	Application	Title
Mouse	WB	Int Orthop; Proteomic analysis of mineralising osteoblasts identifies novel genes related to bone matrix mineralisation.; Authors - Saad Fawzy A FA