Validation Data Gallery

Filter: AllWBIPIHCIF/ICCFC (Intra)

WB analysis using 16675-1-AP
HepG2 lysates were subjected to SDS PAGE followed by western blot with 16675-1-AP (AMFR antibody) at dilution of 1:4000 incubated at 37℃ and 100℃ for 1.5 hours.
WB analysis of HepG2 using 16675-1-AP
WB result of AMFR antibody (16675-1-AP; 1:1000; incubated at room temperature for 1.5 hours) with sh-Control and sh-AMFR transfected HepG2 cells.
WB analysis using 16675-1-AP
Various lysates were subjected to SDS PAGE followed by western blot with 16675-1-AP (AMFR/GP78 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.
IP experiment of MCF-7 using 16675-1-AP
IP result of anti-AMFR/GP78 (IP:16675-1-AP, 4ug; Detection:16675-1-AP 1:300) with MCF-7 cells lysate 2400 ug.
IHC staining of mouse ovary using 16675-1-AP
Immunohistochemical analysis of paraffin-embedded mouse ovary tissue slide using 16675-1-AP (AMFR/GP78 antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of mouse ovary using 16675-1-AP
Immunohistochemical analysis of paraffin-embedded mouse ovary tissue slide using 16675-1-AP (AMFR/GP78 antibody) at dilution of 1:400 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human kidney using 16675-1-AP
Immunohistochemical analysis of paraffin-embedded human kidney tissue slide using 16675-1-AP (AMFR antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human kidney using 16675-1-AP
Immunohistochemical analysis of paraffin-embedded human kidney tissue slide using 16675-1-AP (AMFR antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IF Staining of HepG2 using 16675-1-AP
Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using AMFR/GP78 antibody (16675-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-phalloidin (red).
FC experiment of HepG2 using 16675-1-AP
1X10^6 HepG2 cells were intracellularly stained with 0.4 ug Anti-Human AMFR/GP78 (16675-1-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug rabbit IgG isotype control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer.

Featured Product
KD/KO Validated

AMFR/GP78 Polyclonal antibody

AMFR/GP78 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Cat No. 16675-1-AP

Publications(39)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

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

Synonyms

AMFR, E3 ubiquitin-protein ligase AMFR, EC:2.3.2.36, GP78, RNF 45

Formulation: PBS, Azide, Glycerol

PBS, Azide, Glycerol

Conjugate: Unconjugated

Unconjugated

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (39)
Protocols

データシート［PDF］を印刷

MSDS

Tested Applications

Positive WB detected in	37°C incubated HepG2 cells, HepG2 cells
Positive IP detected in	MCF-7 cells
Positive IHC detected in	mouse ovary tissue, human kidney 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	HepG2 cells

Recommended dilution

Application	Dilution
Western Blot (WB)	WB : 1:1000-1:8000
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) : 0.40 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 9 publications below
WB	See 32 publications below
IHC	See 1 publications below
IF	See 5 publications below
IP	See 1 publications below
CoIP	See 2 publications below

Product Information

16675-1-AP targets AMFR/GP78 in WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA applications and shows reactivity with human, mouse, rat samples.

Tested Reactivity	human, mouse, rat
Cited Reactivity	human, mouse, rat
Host / Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	CatNo: Ag10178 Product name: Recombinant human AMFR protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 301-643 aa of BC069197 Sequence: LFHEVQRRIRRHKNYLRVVGNMEARFAVATPEELAVNNDDCAICWDSMQAARKLPCGHLFHNSCLRSWLEQDTSCPTCRMSLNIADNNRVREEHQGENLDENLVPVAAAEGRPRLNQHNHFFHFDGSRIASWLPSFSVEVMHTTNILGITQASNSQLNAMAHQIQEMFPQVPYHLVLQDLQLTRSVEITTDNILEGRIQVPFPTQRSDSIRPALNSPVERPSSDQEEGETSAQTERVPLDLSPRLEETLDFGEVEVEPSEVEDFEARGSRFSKSADERQRMLVQRKDELLQQARKRFLNKSSEDDAASESFLPSEGASSDPVTLRRRMLAAAAERRLQKQQTS 相同性解析による交差性が予測される生物種
Full Name	autocrine motility factor receptor
Calculated molecular weight	643 aa, 73 kDa
Observed molecular weight	73 kDa
GenBank accession number	BC069197
Gene Symbol	AMFR/GP78
Gene ID (NCBI)	267
RRID	AB_2226463
Conjugate	Unconjugated
Form
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	Q9UKV5
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

AMFR(Autocrine motility factor receptor) is also named as RNF45,gp78.It is the receptor of Autocrine motility factor(AMF),which is a protein secreted by tumor cells that stimulates tumor motility. AMFR has a potential N-glycosylation site, and several potential O-glycosylation sites(PMID:10456327).In normal fibroblasts,the expression of AMFR appeared to be regulated by cell-cell contact(PMID:7626106).

Protocols

Product Specific Protocols
FC protocol for AMFR/GP78 antibody 16675-1-AP	Download protocol
IF protocol for AMFR/GP78 antibody 16675-1-AP	Download protocol
IHC protocol for AMFR/GP78 antibody 16675-1-AP	Download protocol
IP protocol for AMFR/GP78 antibody 16675-1-AP	Download protocol
WB protocol for AMFR/GP78 antibody 16675-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

Species	Application	Title
mouse	WB	Cell BOK Is a Non-canonical BCL-2 Family Effector of Apoptosis Regulated by ER-Associated Degradation. Authors - Fabien Llambi KD Validated View Article
mouse,human	WB,IF	Immunity The E3 Ubiquitin Ligase AMFR and INSIG1 Bridge the Activation of TBK1 Kinase by Modifying the Adaptor STING. Authors - Qiang Wang KD Validated View Article
human	WB	Nat Commun AMFR-mediated Flavivirus NS2A ubiquitination subverts ER-phagy to augment viral pathogenicity Authors - Linliang Zhang View Article
		Cell Death Differ Ubiquitination of NLRP3 by gp78/Insig-1 restrains NLRP3 inflammasome activation. Authors - Ting Xu View Article
human	IF	PLoS Biol Reticulon and CLIMP-63 regulate nanodomain organization of peripheral ER tubules. Authors - Guang Gao View Article
mouse	WB	Brief Bioinform Standigm ASK™: knowledge graph and artificial intelligence platform applied to target discovery in idiopathic pulmonary fibrosis Authors - Seokjin Han View Article

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis using 16675-1-AP

WB analysis of HepG2 using 16675-1-AP

WB analysis using 16675-1-AP

IP experiment of MCF-7 using 16675-1-AP

IHC staining of mouse ovary using 16675-1-AP

IHC staining of mouse ovary using 16675-1-AP

IHC staining of human kidney using 16675-1-AP

IHC staining of human kidney using 16675-1-AP

IF Staining of HepG2 using 16675-1-AP

FC experiment of HepG2 using 16675-1-AP

AMFR/GP78 Polyclonal antibody

AMFR/GP78 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

-/ -

Tested Applications

Recommended dilution

Published Applications

Product Information

Background Information

Protocols

Publications

BOK Is a Non-canonical BCL-2 Family Effector of Apoptosis Regulated by ER-Associated Degradation.

The E3 Ubiquitin Ligase AMFR and INSIG1 Bridge the Activation of TBK1 Kinase by Modifying the Adaptor STING.

AMFR-mediated Flavivirus NS2A ubiquitination subverts ER-phagy to augment viral pathogenicity

Ubiquitination of NLRP3 by gp78/Insig-1 restrains NLRP3 inflammasome activation.

Reticulon and CLIMP-63 regulate nanodomain organization of peripheral ER tubules.

Standigm ASK™: knowledge graph and artificial intelligence platform applied to target discovery in idiopathic pulmonary fibrosis