TOLLIP Monoclonal antibody

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  WB analysis using 68170-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 68170-1-Ig (TOLLIP antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis using 68170-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 68170-1-Ig (TOLLIP antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Beta Actin Monoclonal antibody (HRP-66009) as loading control.

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  WB analysis of HepG2 using 68170-1-Ig

  WB result of TOLLIP antibody (68170-1-Ig; 1:1000; incubated at room temperature for 1.5 hours) with sh-Control and sh-TOLLIP transfected HepG2 cells.

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  IHC staining of mouse colon using 68170-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse colon tissue slide using 68170-1-Ig (TOLLIP antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse colon using 68170-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse colon tissue slide using 68170-1-Ig (TOLLIP antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of rat colon using 68170-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat colon tissue slide using 68170-1-Ig (TOLLIP antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of U2OS using 68170-1-Ig

  Immunofluorescent analysis of (-20°C Methanol) fixed U2OS cells using TOLLIP antibody (68170-1-Ig, Clone: 1B3C4 ) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  FC experiment of Jurkat using 68170-1-Ig

  1X10^6 Jurkat cells were intracellularly stained with 0.4 ug Anti-Human TOLLIP (68170-1-Ig, Clone:1B3C4) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG1 Isotype Control (MOPC-21) (65124-1-Ig, Clone: MOPC-21) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

• Featured Product
• KD/KO Validated

TOLLIP Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Cat No. 68170-1-Ig

Clone No.1B3C4

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat, pig

Applications

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

Synonyms

1B3C4, IL 1RAcPIP, toll interacting protein, Toll-interacting protein

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Tested Applications

• Positive WB detected in: pig brain tissue, LNCaP cells, HepG2 cells, Jurkat cells, K-562 cells, human peripheral blood platelets, rabbit brain tissue, rat brain tissue, mouse brain tissue
• Positive IHC detected in: mouse colon tissue, rat colon 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: U2OS cells
• Positive FC (Intra) detected in: Jurkat cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:400-1:1600
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600
• 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.

Product Information

68170-1-Ig targets TOLLIP in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat, pig samples.

• Tested Reactivity: human, mouse, rat, pig
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag32731

  Product name: Recombinant human TOLLIP protein

  Source: e coli.-derived, PET28a

  Tag: 6*His

  Domain: 1-274 aa of BC012057

  Sequence: MATTVSTQRGPVYIGELPQDFLRITPTQQQRQVQLDAQAAQQLQYGGAVGTVGRLNITVVQAKLAKNYGMTRMDPYCRLRLGYAVYETPTAHNGAKNPRWNKVIHCTVPPGVDSFYLEIFDERAFSMDDRIAWTHITIPESLRQGKVEDKWYSLSGRQGDDKEGMINLVMSYALLPAAMVMPPQPVVLMPTVYQQGVGYVPITGMPAVCSPGMVPVALPPAAVNAQPRCSEEDLKAIQDMFPNMDQEVIRSVLEAQRGNKDAAINSLLQMGEEP

• Full Name: toll interacting protein
• Calculated Molecular Weight: 30 kDa
• Observed Molecular Weight: 30 kDa
• GenBank Accession Number: BC012057
• Gene Symbol: TOLLIP
• Gene ID (NCBI): 54472
• RRID: AB_2923686
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: Q9H0E2
• 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

TOLLIP (Toll interacting protein), is an inhibitory adaptor protein. Tollip is a negative regulator of IL-1R and Toll-like receptors signalling pathway, involved in inflammatory immune response and inhibiting IRAK1 phosphorylation and kinase activity (PMID:11441107, PMID:10854325, PMID:11751856). The TOLLIP-dependent selective autophagy pathway plays a role in clearance of cytotoxic polyQ proteins aggregates (PubMed:25042851).