Invitrogen

SHARPIN Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

View all (19) SHARPIN antibodies

Datasheet

Protocols

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Datasheet

Protocols

Questions & Answers

Tech Support

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SHARPIN Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of SHARPIN was performed using 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with SHARPIN Polyclonal Antibody (Product # PA5-60717) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic(endosomal like pattern) localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

SHARPIN Antibody in Immunohistochemistry (IHC) — Immunohistochemical staining of SHARPIN in human colon tissue shows strong cytoplasmic positivity in glandular cells. Samples were probed using a SHARPIN Polyclonal Antibody (Product # PA5-60717).

SHARPIN Antibody in Western Blot (WB) — Western blot was performed using Anti-SHARPIN Polyclonal Antibody (Product # PA5-60717) and a 42 kDa band corresponding to SHARPIN was observed across cell lines tested. Whole cell extracts (30 µg lysate) of A549 (Lane 1), MCF7 (Lane 2), HeLa (Lane 3), PC-3 (Lane 4), THP-1 (Lane 5) and Jurkat (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.1 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

SHARPIN Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of the target protein. A549 cells were transfected with SHARPIN siRNA and a decrease in signal intensity was observed in Western Blot application using Anti-SHARPIN Polyclonal Antibody (Product # PA5-60717). {KD}

Product Details

PA5-60717

Applications	Tested Dilution	Publications
Western Blot (WB)	0.04-0.4 µg/mL	-
Immunohistochemistry (IHC)	1:20-1:50	-

Product Specifications
Species Reactivity	Human
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant protein corresponding to Human SHARPIN View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.2 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7.2, with 40% glycerol
Contains	0.02% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2647262

Product Specific Information

Immunogen sequence: KSNSPPALGP EACPVSLPSP PEASTLKGPP PEADLPRSPG NLTEREELAG SLARAIAGGD EKGAAQVAAV LAQHRVALSV QLQEACFPPG

Highest antigen sequence identity to the following orthologs: Mouse - 62%, Rat - 57%.

Target Information

Component of the LUBAC complex which conjugates linear polyubiquitin chains in a head-to-tail manner to substrates and plays a key role in NF-kappa-B activation and regulation of inflammation. LUBAC conjugates linear polyubiquitin to IKBKG and RIPK1 and is involved in activation of the canonical NF-kappa-B and the JNK signaling pathways. Linear ubiquitination mediated by the LUBAC complex interferes with TNF-induced cell death and thereby prevents inflammation. LUBAC is proposed to be recruited to the TNF-R1 signaling complex (TNF-RSC) following polyubiquitination of TNF-RSC components by BIRC2 and/or BIRC3 and to conjugate linear polyubiquitin to IKBKG and possibly other components contributing to the stability of the complex. Together with FAM105B/otulin, the LUBAC complex regulates the canonical Wnt signaling during angiogenesis.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: hSIPL1; SHANK-associated RH domain interactor; Shank-associated RH domain-interacting protein; Shank-interacting protein-like 1; Sharpin

Gene Aliases: PSEC0216; SHARPIN; SIPL1

UniProt ID: (Human) Q9H0F6

Entrez Gene ID: (Human) 81858

Molecular Function: ubiquitin-protein ligase protein modifying enzyme

Function(s)

mitochondrion organization I-kappaB kinase/NF-kappaB signaling brain development regulation of tumor necrosis factor-mediated signaling pathway apoptotic nuclear changes keratinization positive regulation of I-kappaB kinase/NF-kappaB signaling proteasome-mediated ubiquitin-dependent protein catabolic process negative regulation of inflammatory response protein homooligomerization protein linear polyubiquitination regulation of CD40 signaling pathway

Process(es)

ubiquitin-protein transferase activity protein binding zinc ion binding polyubiquitin binding protein complex binding identical protein binding

SHARPIN Polyclonal Antibody

Product Details

PA5-60717

Western Blot (WB)

Immunohistochemistry (IHC)

Species Reactivity

Host/Isotype

Class

Type

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

References (0)

Bioinformatics

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