Phospho-RIPK1 (Ser161) Monoclonal antibody

Phospho-RIPK1 (Ser161) Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human and More (2)

Applications

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

Conjugate

Unconjugated

Publications

17

CloneNo.

1B2G1

Cat no : 66854-1-Ig

Synonyms

RIPK1, RIP, Receptor-interacting serine/threonine-protein kinase 1, Receptor-interacting protein 1, Receptor interacting serine/threonine protein kinase 1

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Validation Data Gallery

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  WB analysis using 66854-1-Ig

  Non-treated HEK-293T and Calyculin A treated HEK-293T cells were subjected to SDS PAGE followed by western blot with 66854-1-Ig (Phospho-RIPK1 (Ser161) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

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  WB analysis using 66854-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66854-1-Ig (Phospho-RIPK1 (Ser161) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was untreated (left) or treated with Lambda Protein Phosphatase (λ-PPase, 500U, right) at 37℃ for 1 hours before blocking.

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  IHC staining of human small intestine using 66854-1-Ig

  Immunohistochemical analysis of paraffin-embedded human small intestine tissue slide using 66854-1-Ig (RIPK1 phospho S161 antibody) at dilution of 1:500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human small intestine using 66854-1-Ig

  Immunohistochemical analysis of paraffin-embedded human small intestine tissue slide using 66854-1-Ig (RIPK1 phospho S161 antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human heart using 66854-1-Ig

  Immunohistochemical analysis of paraffin-embedded human heart tissue slide using 66854-1-Ig (RIPK1 phospho S161 antibody) at dilution of 1:500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human heart using 66854-1-Ig

  Immunohistochemical analysis of paraffin-embedded human heart tissue slide using 66854-1-Ig (RIPK1 phospho S161 antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of U2OS using 66854-1-Ig

  Immunofluorescent analysis of (-20°C Ethanol) fixed U2OS cells using Phospho-RIPK1 (Ser161) antibody (66854-1-Ig, Clone: 1B2G1 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1).

• 

  FC experiment of HEK-293T using 66854-1-Ig

  1X10^6 Calyculin A treated HEK-293T cells were intracellularly stained with 0.13 ug Anti-Human Phospho-RIPK1 (Ser161) (66854-1-Ig, Clone:1B2G1) labeled with FlexAble CoraLite® Plus 555 Antibody Labeling Kit for Mouse IgG1 (KFA022). Cells were fixed with 4% PFA and permeabilized with 90% MeOH.

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Tested Applications

• Positive WB detected in: Jurkat cells, Calyculin A treated HEK-293T cells, Raji cells, THP-1 cells
• Positive IHC detected in: human small intestine tissue, human heart 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: Calyculin A treated HEK-293T cells

Recommended dilution

• Western Blot (WB): WB : 1:2000- 1:20000
• Immunohistochemistry (IHC): IHC : 1:250-1:1000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.13 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 16 publications below
• IHC: See 4 publications below
• IF: See 2 publications below

Product Information

66854-1-Ig targets Phospho-RIPK1 (Ser161) in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: receptor (TNFRSF)-interacting serine-threonine kinase 1
• Calculated Molecular Weight: 76 kDa
• Observed Molecular Weight: 74 kDa, 45 kDa
• GenBank Accession Number: NM_003804
• Gene Symbol: RIPK1
• Gene ID (NCBI): 8737
• RRID: AB_2882194
• 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

RIPK1(Receptor-interacting serine/threonine-protein kinase 1) is primarily involved in mediating TNF-R1-induced cell activation, apoptosis and necroptosis and belongs to a novel class of kinases thatfunction in cell survival and cell death mechanisms(PMID:22685397 ).It has 2 isoforms produced by alternative splicing.It also can exist as the protein with lower molecular weight about 25kd and 45kd in HAEC and HUVEC(PMID:22685397).

Publications

Species	Application	Title
human	WB	Cancer Lett; NR4A1 depletion inhibits colorectal cancer progression by promoting necroptosis via the RIG-I-like receptor pathway; Authors - Jinghan Zhu
mouse	WB,IHC,IF	Cell Commun Signal; IRF1 regulation of ZBP1 links mitochondrial DNA and chondrocyte damage in osteoarthritis; Authors - Kai Sun
human	WB	Inflammation; Ursolic Acid Promotes Autophagy by Inhibiting Akt/mTOR and TNF-α/TNFR1 Signaling Pathways to Alleviate Pyroptosis and Necroptosis in Mycobacterium tuberculosis-Infected Macrophages; Authors - Jingjing Shen
human	WB	J Cell Mol Med; LRG-1 promotes fat graft survival through the RAB31-mediated inhibition of hypoxia-induced apoptosis.; Authors - Chia-Kang Ho
mouse	WB	BMC Complement Med Ther; JianPiYiShen formula prevents cisplatin-induced acute kidney injury in mice by improving necroptosis through MAPK pathway; Authors - Zhongtang Li
human	WB	Front Genet; Exosome-Derived LncRNA TCONS_00072128 Mediated Osteogenic Differentiation and Inflammation by Caspase 8 Regulation.; Authors - Yongchang Yang