Validation Data Gallery

Filter: AllWBIPIHCIF-PIF/ICCFC (Intra)

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.
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.
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.
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.
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.
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).
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).
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).
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).
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 isotype control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer.

Nucleobindin 1 Polyclonal antibody

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

Cat No. 10228-1-AP

Publications(1)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

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

Synonyms

NUCB1, CALNUC, NUC, Nucleobindin-1

Formulation: PBS, Azide, Glycerol

PBS, Azide, Glycerol

Conjugate: Unconjugated

Unconjugated

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (1)
Protocols

データシート［PDF］を印刷

MSDS

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

Application	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	CatNo: Ag0290 Product name: Recombinant human NUCB1 protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 127-286 aa of BC002356 Sequence: VQVDHLNLLKQFEHLDPQNQHTFEARDLELLIQTATRDLAQYDAAHHEEFKRYEMLKEHERRRYLESLGEEQRKEAERKLEEQQRRHREHPKVNVPGSQAQLKEVWEELDGLDPNRFNPKTFFILHDINSDGVLDEQELEALFTKELEKVYDPKNEEDDM 相同性解析による交差性が予測される生物種
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
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	Q02818
Storage Buffer	PBS with 0.02% sodium azide and 50% glycerol{{ptg:BufferTemp}}7.3
Storage Conditions	Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

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)

Protocols

Product Specific Protocols
IF protocol for Nucleobindin 1 antibody 10228-1-AP	Download protocol
IHC protocol for Nucleobindin 1 antibody 10228-1-AP	Download protocol
IP protocol for Nucleobindin 1 antibody 10228-1-AP	Download protocol
WB protocol for Nucleobindin 1 antibody 10228-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

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 View Article

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

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

WB analysis of HeLa using 10228-1-AP

WB analysis of A431 using 10228-1-AP

WB analysis of HepG2 using 10228-1-AP

IP experiment of HepG2 using 10228-1-AP

IHC staining of human brain using 10228-1-AP