ECSIT Recombinant antibody

ECSIT Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

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

Conjugate

Unconjugated

CloneNo.

240223G2

Cat no : 83295-5-RR

Synonyms

SITPEC, Protein SITPEC, ECSIT homolog (Drosophila), 240223G2

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Validation Data Gallery

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  WB analysis using 83295-5-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 83295-5-RR (ECSIT antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IHC staining of human intrahepatic cholangiocarcinoma using 83295-5-RR

  Immunohistochemical analysis of paraffin-embedded human intrahepatic cholangiocarcinoma tissue slide using 83295-5-RR (ECSIT antibody) at dilution of 1:250 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human intrahepatic cholangiocarcinoma using 83295-5-RR

  Immunohistochemical analysis of paraffin-embedded human intrahepatic cholangiocarcinoma tissue slide using 83295-5-RR (ECSIT antibody) at dilution of 1:250 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of A431 using 83295-5-RR

  Immunofluorescent analysis of (4% PFA) fixed A431 cells using ECSIT antibody (83295-5-RR, Clone: 240223G2 ) at dilution of 1:200 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002), CL594-phalloidin (red).

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  FC experiment of HeLa using 83295-5-RR

  1x10^6 HeLa cells were intracellularly stained with 0.25 ug ECSIT Recombinant antibody (83295-5-RR, Clone:240223G2) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug Rabbit IgG Isotype Control Recombinant Antibody (98136-1-RR, Clone: 240953C9) (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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  Affinity and Kinetic Characterization of 83295-5-RR

  Biolayer interferometry (BLl) kinetic assays of 83295-5-RR against Human ECSIT were performed. The affinity constant is 1.95 nM.

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Tested Applications

• Positive WB detected in: HeLa cells, Jurkat cells, K-562 cells, A549 cells, HepG2 cells, HEK-293 cells
• Positive IHC detected in: human intrahepatic cholangiocarcinoma 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: A431 cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:125-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 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

The immunogen of 83295-5-RR is ECSIT Fusion Protein expressed in E. coli.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: ECSIT fusion protein Ag26898
• Full Name: ECSIT homolog (Drosophila)
• Calculated Molecular Weight: 49 kDa
• Observed Molecular Weight: 49 kDa
• GenBank Accession Number: BC008279
• Gene Symbol: ECSIT
• Gene ID (NCBI): 51295
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• 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

ECSIT is a cytosolic adaptor protein associated with the toll-like receptor pathway. It has a distinct N-terminal mitochondrial targeting sequence, pentatricopeptide repeat motif, and a C-terminal pleckstrin homology domain. ECSIT regulates many biological processes like embryonic development, inflammation, cardiac function, and assembly of mitochondrial complex I. In macrophages, the activation of TLRs by inflammatory signals promotes the translocation of TRAF6 to mitochondria where it engages ECSIT to stimulate the production of mitochondrial reactive oxygen species (ROS).