USP4 Monoclonal antibody

USP4 Monoclonal Antibody for WB, IHC, IF-P, FC (Intra), ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

Human, Mouse and More (2)

Applications

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

Conjugate

Unconjugated

Publications

7

CloneNo.

1G11G8

Cat no : 66822-1-Ig

Synonyms

Unph, UNP, Ubiquitin-specific-processing protease 4, Ubiquitin carboxyl-terminal hydrolase 4, EC:3.4.19.12

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Validation Data Gallery

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  WB analysis using 66822-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66822-1-Ig (USP4 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis using 66822-1-Ig

  HEK-293 cells and Jurkat cells were subjected to SDS PAGE followed by western blot with 66822-1-Ig (USP4 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  IHC staining of human breast cancer using 66822-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66822-1-Ig (USP4 antibody) at dilution of 1:300 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 66822-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66822-1-Ig (USP4 antibody) at dilution of 1:300 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse kidney using 66822-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 66822-1-Ig (USP4 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse kidney using 66822-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 66822-1-Ig (USP4 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of human breast cancer using 66822-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed human breast cancer tissue using USP4 antibody (66822-1-Ig, Clone: 1G11G8 ) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

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  IF Staining of human liver cancer using 66822-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed human liver cancer tissue using USP4 antibody (66822-1-Ig, Clone: 1G11G8 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

• 

  FC experiment of THP-1 using 66822-1-Ig

  1X10^6 THP-1 cells were intracellularly stained with 0.4 ug Anti-Human USP4 (66822-1-Ig, Clone:1G11G8) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG2b isotype control Mouse McAb (66360-3-Ig, Clone: 11B8C4) (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: HeLa cells, Jurkat cells, HEK-293 cells, THP-1 cells, NCI-H1299 cells, RAW 264.7 cells
• Positive IHC detected in: human breast cancer tissue, mouse 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-P detected in: human breast cancer tissue, human liver cancer tissue
• Positive FC (Intra) detected in: THP-1 cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:6000
• Immunohistochemistry (IHC): IHC : 1:100-1:300
• Immunofluorescence (IF)-P: IF-P : 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

• WB: See 6 publications below
• IHC: See 2 publications below
• IF: See 1 publications below
• IP: See 1 publications below

Product Information

66822-1-Ig targets USP4 in WB, IHC, IF-P, FC (Intra), IP, ELISA applications and shows reactivity with Human, Mouse samples.

• Tested Reactivity: Human, Mouse
• Cited Reactivity: human, mouse
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: USP4 fusion protein Ag28297
• Full Name: ubiquitin specific peptidase 4 (proto-oncogene)
• Calculated Molecular Weight: 963 aa, 109 kDa
• Observed Molecular Weight: 109 kDa
• GenBank Accession Number: BC125131
• Gene Symbol: USP4
• Gene ID (NCBI): 7375
• RRID: AB_2882165
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A 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

USP4, also named as UNP and UNPH, belongs to the peptidase C19 family and USP4 subfamily. USP4 is a deubiquitinating enzyme that links to mitogen-activated protein kinase signaling, pre-mRNA splicing, and control of p53 stability (PMID: 26455393). USP4 has 3 isoforms with the molecular mass of 36, 104 and 109 kDa. Recently, it has been reported that USP4 is a critical factor in promoting lung cancer stemness and potentially useful lung cancer prognosis marker (PMID: 32549341).

Publications

Species	Application	Title
human	IHC	Cell Death Dis; USP4 promotes the proliferation, migration, and invasion of esophageal squamous cell carcinoma by targeting TAK1; Authors - Hongdian Zhang
mouse	WB	bioRxiv; The DNA helicase FANCJ (BRIP1) functions in Double Strand Break repair processing, but not crossover formation during Prophase I of meiosis in male mice; Authors - Tegan S Horan
human	WB,IF,IP	PLoS One; USP4 promotes the proliferation and glucose metabolism of gastric cancer cells by upregulating PKM2; Authors - Yuanyuan Chen
mouse	WB	PLoS Genet; The DNA helicase FANCJ (BRIP1) functions in double strand break repair processing, but not crossover formation during prophase I of meiosis in male mice; Authors - Tegan S Horan
human	WB	Cell Rep; Oncogenic KRAS effector USP13 promotes metastasis in non-small cell lung cancer through deubiquitinating β-catenin; Authors - Yanguan Guo
human	WB,IHC	NPJ Breast Cancer; The deubiquitinating enzyme USP4 regulates BRCA1 stability and function; Authors - Xueyuan Guo