SHARPIN Polyclonal antibody

• Featured Product
• KD/KO Validated

SHARPIN Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse and More (1)

Applications

WB, IHC, IF/ICC, IP, CoIP, ELISA

Conjugate

Unconjugated

Publications

103

Reviews (1)

Cat no : 14626-1-AP

Synonyms

SIPL1, Shank-interacting protein-like 1, Shank-associated RH domain-interacting protein, PSEC0216, hSIPL1

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Validation Data Gallery

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  WB analysis using 14626-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 14626-1-AP (SHARPIN antibody) at dilution of 1:40000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of Jurkat using 14626-1-AP

  Jurkat cells were subjected to SDS PAGE followed by western blot with 14626-1-AP (SHARPIN Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of A549 using 14626-1-AP

  A549 cells were subjected to SDS PAGE followed by western blot with 14626-1-AP (SHARPIN antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse brain using 14626-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 14626-1-AP (SHARPIN Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse skeletal muscle using 14626-1-AP

  mouse skeletal muscle tissue were subjected to SDS PAGE followed by western blot with 14626-1-AP (SHARPIN Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  IP experiment of Raji using 14626-1-AP

  IP result of anti-SHARPIN (IP:14626-1-AP, 3ug; Detection:14626-1-AP 1:1500) with Raji cells lysate 2520ug.

• 

  IHC staining of human lymphoma using 14626-1-AP

  Immunohistochemical analysis of paraffin-embedded human lymphoma tissue slide using 14626-1-AP (SHARPIN Antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lymphoma using 14626-1-AP

  Immunohistochemical analysis of paraffin-embedded human lymphoma tissue slide using 14626-1-AP (SHARPIN Antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human small intestine using 14626-1-AP

  Immunohistochemical analysis of paraffin-embedded human small intestine tissue slide using 14626-1-AP (SHARPIN Antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human small intestine using 14626-1-AP

  Immunohistochemical analysis of paraffin-embedded human small intestine tissue slide using 14626-1-AP (SHARPIN Antibody) at dilution of 1:50 (under 40x lens).

• 

  IF Staining of Jurkat using 14626-1-AP

  Immunofluorescent analysis of (4% PFA) fixed Jurkat cells using SHARPIN antibody (14626-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

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Tested Applications

• Positive WB detected in: A549 cells, mouse brain tissue, mouse skeletal muscle tissue, Jurkat cells, Raji cells
• Positive IP detected in: Raji cells
• Positive IHC detected in: human lymphoma tissue, human small intestine 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: Jurkat cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:100-1:400
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500

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 12 publications below
• WB: See 94 publications below
• IHC: See 2 publications below
• IF: See 3 publications below
• IP: See 16 publications below
• CoIP: See 3 publications below

Product Information

14626-1-AP targets SHARPIN in WB, IHC, IF/ICC, IP, CoIP, ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: SHARPIN fusion protein Ag6201
• Full Name: SHANK-associated RH domain interactor
• Calculated Molecular Weight: 40 kDa
• Observed Molecular Weight: 40 kDa
• GenBank Accession Number: BC025244
• Gene Symbol: SHARPIN
• Gene ID (NCBI): 81858
• RRID: AB_2187734
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity 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

SHARPIN is as a third component of the linear ubiquitin chain assembly complex (LUBAC), recruited to the CD40 and TNF receptor signaling complexes together with its other constituents, HOIL1 and HOIP(PMID:21455173).SHARPIN is enriched in the postsynaptic density and forms a complex with SHANK in heterologous cells and brain(PMID:11178875).It has 2 isoforms produced by alternative splicing.

Publications

Species	Application	Title
mouse	WB	Nature; OTULIN limits cell death and inflammation by deubiquitinating LUBAC.; Authors - Klaus Heger
mouse	WB,IP	Nature; LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis.; Authors - Nieves Peltzer
human	WB,IP	Cell; OTULIN Antagonizes LUBAC Signaling by Specifically Hydrolyzing Met1-Linked Polyubiquitin.; Authors - Keusekotten Kirstin K
human	WB	Cell; The Deubiquitinase OTULIN Is an Essential Negative Regulator of Inflammation and Autoimmunity.; Authors - Rune Busk Damgaard
mouse	WB	Cancer Cell; A Dual Role of Caspase-8 in Triggering and Sensing Proliferation-Associated DNA Damage, a Key Determinant of Liver Cancer Development.; Authors - Yannick Boege
mouse	WB	Nat Immunol; Pellino3 ubiquitinates RIP2 and mediates Nod2-induced signaling and protective effects in colitis.; Authors - Yang Shuo S

Reviews

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

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FH

Katharina (Verified Customer) (07-17-2019)

The antibody worked as expected, detecting a nice band at the right size. Recommended

• Applications: Western Blot,
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: A549 cells /immunoprecipitates

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