AMFR/GP78 Polyclonal antibody

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  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.

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  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.

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  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.

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  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.

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  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).

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  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).

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  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).

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  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).

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  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).

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  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 (PF00011-C).

• Featured Product
• KD/KO Validated

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

Cat No. 16675-1-AP

Publications(38)

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

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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

• 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 31 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

• 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: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: Q9UKV5
• 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

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).

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

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
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
human	WB	Nat Commun; AMFR-mediated Flavivirus NS2A ubiquitination subverts ER-phagy to augment viral pathogenicity; Authors - Linliang Zhang
		Cell Death Differ; Ubiquitination of NLRP3 by gp78/Insig-1 restrains NLRP3 inflammasome activation.; Authors - Ting Xu
human	IF	PLoS Biol; Reticulon and CLIMP-63 regulate nanodomain organization of peripheral ER tubules.; Authors - Guang Gao
mouse	WB	Brief Bioinform; Standigm ASK™: knowledge graph and artificial intelligence platform applied to target discovery in idiopathic pulmonary fibrosis; Authors - Seokjin Han