Filter: AllWBIPIHCIF/ICCFC (Intra)

WB analysis using 20175-1-AP
Various lysates were subjected to SDS PAGE followed by western blot with 20175-1-AP (NCOA5 antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.
IP experiment of HEK-293 using 20175-1-AP
IP result of anti-NCOA5 (IP:20175-1-AP, 4ug; Detection:20175-1-AP 1:800) with HEK-293 cells lysate 3000ug.
IHC staining of human stomach using 20175-1-AP
Immunohistochemical analysis of paraffin-embedded human stomach tissue slide using 20175-1-AP (NCOA5 antibody) at dilution of 1:400 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IF Staining of A431 using 20175-1-AP
Immunofluorescent analysis of (4% PFA) fixed A431 cells using NCOA5 antibody (20175-1-AP) at dilution of 1:200 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002), CL594-phalloidin (red).
FC experiment of HEK-293 using 20175-1-AP
1x10^6 HEK-293 cells were intracellularly stained with 0.25 ug NCOA5 Polyclonal antibody (20175-1-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 and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

Featured Product
KD/KO Validated

NCOA5 Polyclonal antibody

NCOA5 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Cat No. 20175-1-AP

Publications(3)

Reviews (5)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (1)

Applications

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

Synonyms

CIA, Coactivator independent of AF-2, KIAA1637, NCoA-5, Nuclear receptor coactivator 5

Formulation: PBS, Azide, Glycerol

PBS, Azide, Glycerol

Conjugate: Unconjugated

Unconjugated

Size/Concentration:

-/ -

Freight/Packing: -

Quantity

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Overview

Tested Applications
Product Information
Publications (3)
Protocols
Reviews (5)

Print datasheet

MSDS SDS

Tested Applications

Positive WB detected in	HEK-293 cells, HeLa cells
Positive IP detected in	HEK-293 cells
Positive IHC detected in	human stomach 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	A431 cells
Positive FC (Intra) detected in	HEK-293 cells

Recommended dilution

Application	Dilution
Western Blot (WB)	WB : 1:1000-1:4000
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 : 1:50-1:500
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

KD/KO	See 2 publications below
WB	See 2 publications below
IHC	See 1 publications below
IF	See 2 publications below
ChIP	See 1 publications below

Product Information

20175-1-AP targets NCOA5 in WB, IHC, IF/ICC, FC (Intra), IP, ChIP, ELISA applications and shows reactivity with human, mouse, rat samples.

Tested Reactivity	human, mouse, rat
Cited Reactivity	human, mouse, bovine
Host / Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	CatNo: Ag14014 Product name: Recombinant human NCOA5 protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 112-384 aa of BC056872 Sequence: DMRDSRDPMYRREGSYDRYLRMDDYCRRKDDSYFDRYRDSFDGRGPPGPESQSRAKERLKREERRREELYRQYFEEIQRRFDAERPVDCSVIVVNKQTKDYAESVGRKVRDLGMVVDLIFLNTEVSLSQALEDVSRGGSPFAIVITQQHQIHRSCTVNIMFGTPQEHRNMPQADAMVLVARNYERYKNECREKEREEIARQAAKMADEAILQERERGGPEEGVRGGHPPAIQSLINLLADNRYLTAEETDKIINYLRERKERLMRSSTDSLPG Predict reactive species
Full Name	nuclear receptor coactivator 5
Calculated Molecular Weight	579 aa, 66 kDa
Observed Molecular Weight	66 kDa
GenBank Accession Number	BC056872
Gene Symbol	NCOA5
Gene ID (NCBI)	57727
RRID	AB_10694674
Conjugate	Unconjugated
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	Q9HCD5
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

Nuclear receptor coactivator 5 (NCOA5), also known as coactivator independent of AF-2 (CIA), possesses both repressor and activator functions (PMID: 11113208). It can interact with Estrogen Receptor (ER) alpha and ER beta in a ligand-dependent manner. NCOA5 has been shown to regulate the ER alpha-mediated transcription as well as the expression of the c-Myc gene (PMID: 11113208; 15073177). Recently identified as a conserved component of pluripotent stem cells, mammalian NCOA5 may contribute to the maintenance of the pluripotent stem cell population (PMID: 24268775).

Protocols

Product Specific Protocols
IF protocol for NCOA5 antibody 20175-1-AP	Download protocol
IHC protocol for NCOA5 antibody 20175-1-AP	Download protocol
IP protocol for NCOA5 antibody 20175-1-AP	Download protocol
WB protocol for NCOA5 antibody 20175-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

Species	Application	Title
mouse	IF	Cell Rep SILAC Proteomics of Planarians Identifies Ncoa5 as a Conserved Component of Pluripotent Stem Cells. Authors - B?ser Alexander A View Article
bovine	WB,IF,ChIP	Biochem Biophys Res Commun NCOA5 is a master regulator of amino acid-induced mTOR activation and β-casein synthesis in bovine mammary epithelial cells. Authors - Xiaohan Yuan KD Validated View Article
human	WB,IHC	Cell Death Discov NCOA5 induces sorafenib resistance in hepatocellular carcinoma by inhibiting ferroptosis Authors - Shuang Gao KD Validated View Article

Reviews

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

FH Matthew (Verified Customer) (10-01-2025)	Got great signal on both my western blot and immunofluorescence experiments when over-expressing NCOA5. I will definitely use in the future for other applications! Applications: Western Blot, Immunofluorescence Primary Antibody Dilution: 1:100 (IF); 1:1000 (WB) Cell Tissue Type: 22rv1, PC3
FH Moritz (Verified Customer) (09-26-2025)	worked well for HEK293T cells Applications: Western Blot Primary Antibody Dilution: 1:2000 Cell Tissue Type: HEK293T NCOA5 Antibody Western Blot validation (1:2000 dilution) in HEK293T (Cat no:20175-1-AP)
FH Aleksandr (Verified Customer) (12-16-2024)	Excellent antibody Applications: Western Blot Primary Antibody Dilution: 1:2500 Cell Tissue Type: RKO NCOA5 Antibody Western Blot validation (1:2500 dilution) in RKO (Cat no:20175-1-AP)
FH Tsimafei (Verified Customer) (08-07-2024)	Antibody recognize Ncoa5 band as well as (weakly) the upper non-specific band Applications: Western Blot Primary Antibody Dilution: 1:2000 Cell Tissue Type: U2OS NCOA5 Antibody Western Blot validation (1:2000 dilution) in U2OS (Cat no:20175-1-AP)
FH Julia (Verified Customer) (07-16-2024)	Alright. You can see some non-specific bands and ladder-binding but the correct band is more distinct. Applications: Western Blot Primary Antibody Dilution: 1:2000 Cell Tissue Type: mESC NCOA5 Antibody Western Blot validation (1:2000 dilution) in mESC (Cat no:20175-1-AP)