Synthetic peptide within Human Id1 aa 11-60 / 155.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, FC, mIHC
Target Molecular Weight
Predicted band size: 16 kDa
Positive Control
293T cell lysates, Hela cell lysate, HepG2 cell lysate, A549, Hela, HepG2, human tonsil tissue, human liver tissue, human colon tissue, human placenta tissue.
DNA-binding protein inhibitor ID-1 is a protein that in humans is encoded by the ID1 gene. The protein encoded by this gene is a helix-loop-helix (HLH) protein that can form heterodimers with members of the basic HLH family of transcription factors. The encoded protein has no DNA binding activity and therefore can inhibit the DNA binding and transcriptional activation ability of basic HLH proteins with which it interacts. This protein may play a role in cell growth, senescence, and differentiation. Two transcript variants encoding different isoforms have been found for this gene. ID1 has been shown to interact weakly with MyoD but very tightly with ubiquitously expressed E proteins. E proteins heterodimerize with tissue restricted bHLH proteins such as Myod, NeuroD, etc. to form active transcription complexes so by sequestering E proteins, Id proteins can inhibit tissue restricted gene expression in multiple cell lineages using the same biochemical mechanism. Other interacting partners include CASK. ID1 can be used to mark endothelial progenitor cells which are critical to tumor growth and angiogenesis. Targeting ID1 results in decreased tumor growth. ID1 has been shown to be targeted by cannabidiol in certain gliomas and breast cancers.
Background References
1. Montalbano M et al. Modeling of Hepatocytes Proliferation Isolated from Proximal and Distal Zones from Human Hepatocellular Carcinoma Lesion. PLoS One 11:e0153613 (2016).
2. Li J et al. ATF3 suppresses ESCC via downregulation of ID1. Oncol Lett 12:1642-1648 (2016).
Western blot analysis of Id1 on 293T cell lysates with Rabbit anti-Id1 antibody (ET1705-49) at 1/500 dilution.
Lysates/proteins at 10 µg/Lane.
Predicted band size: 16 kDa Observed band size: 22 kDa
Exposure time: 2 minutes;
15% 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 (ET1705-49) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
Western blot analysis of Id1 on different lysates with Rabbit anti-Id1 antibody (ET1705-49) at 1/500 dilution.
Lane 1: Hela cell lysate Lane 2: HepG2 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 16 kDa Observed band size: 22 kDa
Exposure time: 1 minute;
15% 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 (ET1705-49) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
ICC staining Id1 in A549 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Immunocytochemistry analysis of HeLa cells labeling Id1 with Rabbit anti-Id1 antibody (ET1705-49) 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-Id1 antibody (ET1705-49) 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.
ICC staining Id1 in HepG2 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Id1 antibody. Counter stained with hematoxylin.
Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-Id1 antibody. Counter stained with hematoxylin.
Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-Id1 antibody. Counter stained with hematoxylin.
Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-Id1 antibody. Counter stained with hematoxylin.
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