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SIAH1 Recombinant Rabbit Monoclonal Antibody [PSH01-80]

Usd: 385

Specification

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Catalog# HA721732

SIAH1 Recombinant Rabbit Monoclonal Antibody [PSH01-80]

Application

  • WB

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

SIAH1 Recombinant Rabbit Monoclonal Antibody [PSH01-80]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human SIAH1 aa 1-50 / 282.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell

Molecular Weight

Predicted band size: 31 kDa

Positive Control

HCT 116 cell lysate, RAW264.7 cell lysate, mouse liver tissue lysate, mouse brain tissue lysate, rat liver tissue lysate, rat brain tissue lysate, Rat testis testis lysate, A549.

Conjugation

unconjugated

Clone Number

PSH01-80

RRID

AB_3072844

Reactivity Data

WB IF-Cell
Human
Tested Tested
Tested Tested
Mouse
Tested Tested
Rat
Tested Tested

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

Target

Function

E3 ubiquitin-protein ligase that mediates ubiquitination and subsequent proteasomal degradation of target proteins. E3 ubiquitin ligases accept ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then directly transfers the ubiquitin to targeted substrates.Mediates E3 ubiquitin ligase activity either through direct binding to substrates or by functioning as the essential RING domain subunit of larger E3 complexes.Triggers the ubiquitin-mediated degradation of many substrates, including proteins involved in transcription regulation (ELL2, MYB, POU2AF1, PML and RBBP8), a cell surface receptor (DCC), the cell-surface receptor-type tyrosine kinase FLT3, the cytoplasmic signal transduction molecules (KLF10/TIEG1 and NUMB), an antiapoptotic protein (BAG1), a microtubule motor protein (KIF22), a protein involved in synaptic vesicle function in neurons (SYP), a structural protein (CTNNB1) and SNCAIP. Confers constitutive instability to HIPK2 through proteasomal degradation. It is thereby involved in many cellular processes such as apoptosis, tumor suppression, cell cycle, axon guidance, transcription regulation, spermatogenesis and TNF-alpha signaling. Has some overlapping function with SIAH2 .Induces apoptosis in cooperation with PEG3 (By similarity). Upon nitric oxid (NO) generation that follows apoptotic stimulation, interacts with S-nitrosylated GAPDH, mediating the translocation of GAPDH to the nucleus (By similarity). GAPDH acts as a stabilizer of SIAH1, facilitating the degradation of nuclear proteins (By similarity). Mediates ubiquitination and degradation of EGLN2 and EGLN3 in response to the unfolded protein response (UPR), leading to their degradation and subsequent stabilization of ATF4 (By similarity). Also part of the Wnt signaling pathway in which it mediates the Wnt-induced ubiquitin-mediated proteasomal degradation of AXIN1

Background References

1. Zhang H, Wang J, Ge Y, Ye M, Jin X. Siah1 in cancer and nervous system diseases (Review). Oncol Rep. 2022 Feb;47(2):35. doi: 10.3892/or.2021.8246. Epub 2021 Dec 27. PMID: 34958110.

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: Q8IUQ4 Human

SwissProt: P61092 Mouse

SwissProt: Q920M9 Rat

Synonyms

E3 ubiquitin-protein ligase SIAH1 antibody

FLJ08065 antibody

hSIAH1 antibody

HUMSIAH antibody

Seven in absentia homolog 1 (Drosophila) antibody

Seven in absentia homolog 1 antibody

Siah 1 antibody

Siah 1a antibody

Siah E3 ubiquitin protein ligase 1 antibody

Siah-1 antibody

Expand

Images

  • Western blot analysis of SIAH1 on different lysates with Rabbit anti-SIAH1 antibody (<a href="/products/HA721732" style="font-weight: bold;text-decoration: underline;">HA721732</a>) at 1/1,000 dilution.<br /><br />Lane 1: HCT 116 cell lysate（20 µg/Lane）<br />Lane 2: RAW264.7 cell lysate（20 µg/Lane）<br />Lane 3: Mouse liver tissue lysate（40 µg/Lane）<br />Lane 4: Mouse brain tissue lysate（40 µg/Lane）<br />Lane 5: Rat liver tissue lysate（40 µg/Lane）<br />Lane 6: Rat brain tissue lysate（40 µg/Lane）<br />Lane 7: Rat testis testis lysate（40 µg/Lane）<br /><br />Predicted band size: 31 kDa<br />Observed band size: 25 kDa<br /><br />Exposure time: 20 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721732" style="font-weight: bold;text-decoration: underline;">HA721732</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of SIAH1 on different lysates with Rabbit anti-SIAH1 antibody (HA721732) at 1/1,000 dilution.

    Lane 1: HCT 116 cell lysate（20 µg/Lane）
    Lane 2: RAW264.7 cell lysate（20 µg/Lane）
    Lane 3: Mouse liver tissue lysate（40 µg/Lane）
    Lane 4: Mouse brain tissue lysate（40 µg/Lane）
    Lane 5: Rat liver tissue lysate（40 µg/Lane）
    Lane 6: Rat brain tissue lysate（40 µg/Lane）
    Lane 7: Rat testis testis lysate（40 µg/Lane）

    Predicted band size: 31 kDa
    Observed band size: 25 kDa

    Exposure time: 20 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721732) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of SIAH1 on different lysates with Rabbit anti-SIAH1 antibody (<a href="/products/HA721732" style="font-weight: bold;text-decoration: underline;">HA721732</a>) at 1/1,000 dilution.<br /><br />Lane 1: Jurkat cell lysate<br />Lane 2: HeLa cell lysate<br />Lane 3: A549 cell lysate<br />Lane 4: HCT116 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 31 kDa<br />Observed band size: 31/25kDa<br /><br />Exposure time: 43 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721732" style="font-weight: bold;text-decoration: underline;">HA721732</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of SIAH1 on different lysates with Rabbit anti-SIAH1 antibody (HA721732) at 1/1,000 dilution.

    Lane 1: Jurkat cell lysate
    Lane 2: HeLa cell lysate
    Lane 3: A549 cell lysate
    Lane 4: HCT116 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 31 kDa
    Observed band size: 31/25kDa

    Exposure time: 43 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721732) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of A549 cells labeling SIAH1 with Rabbit anti-SIAH1 antibody (<a href="/products/HA721732" style="font-weight: bold;text-decoration: underline;">HA721732</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SIAH1 antibody (<a href="/products/HA721732" style="font-weight: bold;text-decoration: underline;">HA721732</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of A549 cells labeling SIAH1 with Rabbit anti-SIAH1 antibody (HA721732) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SIAH1 antibody (HA721732) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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