SIAH1 Recombinant antibody

SIAH1 Uni-rAb^TM Recombinant Antibody for WB, IHC, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IHC, ELISA

Conjugate

Unconjugated

CloneNo.

240377B4

Cat no : 83389-4-RR

Synonyms

Siah-1a, Siah-1, RING-type E3 ubiquitin transferase SIAH1, EC:2.3.2.27, E3 ubiquitin-protein ligase SIAH1

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Validation Data Gallery

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  WB analysis using 83389-4-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 83389-4-RR (SIAH1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse kidney using 83389-4-RR

  mouse kidney tissue were subjected to SDS PAGE followed by western blot with 83389-4-RR (SIAH1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  IHC staining of mouse testis using 83389-4-RR

  Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 83389-4-RR (SIAH1 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  FC experiment of A549 using 83389-4-RR

  1x10^6 A549 cells were intracellularly stained with 0.25 ug Siah1 Recombinant Antibody (83389-4-RR, Clone:240377B4) and APC-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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  Affinity and Kinetic Characterization of 83389-4-RR

  Biolayer interferometry (BLl) kinetic assays of 83389-4-RR against Human SIAH1 were performed. The affinity constant is 2.39 nM.

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Tested Applications

• Positive WB detected in: A549 cells, mouse kidney tissue, HepG2 cells, HEK-293T cells, SH-SY5Y cells, Jurkat cells, COLO 320 cells
• Positive IHC detected in: mouse testis tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive FC (Intra) detected in: A549 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunohistochemistry (IHC): IHC : 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 83389-4-RR is SIAH1 Fusion Protein expressed in E. coli.

• Tested Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: SIAH1 fusion protein Ag34618
• Full Name: seven in absentia homolog 1 (Drosophila)
• Calculated Molecular Weight: 282 aa, 31 kDa
• Observed Molecular Weight: 30-34 kDa
• GenBank Accession Number: BC035562
• Gene Symbol: SIAH1
• Gene ID (NCBI): 6477
• 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

SIAH1, also named as HUMSIAH, Siah-1a and Siah-1S(21kd), belongs to the SINA (Seven in absentia) family. SIAH1 is known to cause indirect degradation of beta-catenin through formation of a complex with Siah-interacting protein (SIP), Skp1 and Ebi. SIAH1 can form heterodimer or homodimer such as Siah-1*Siah-1(70 or 62kd), Siah-1*Siah-1S(42kd) and Siah-1S*Siah-1S(51-56kd). Importantly, results from in vitro soft agar assay demonstrated that Siah-1S displays a promotion effect on cells tumorigenicity. (PMID:17420721)