SET Recombinant Rabbit Monoclonal Antibody [JB60-34]
Catalog# ET7107-18
SET Recombinant Rabbit Monoclonal Antibody [JB60-34]
-
WB
-
IF-Cell
-
IHC-P
-
IP
-
IF-Tissue
-
Human
-
Mouse
-
unconjugated
Overview
Product Name
SET Recombinant Rabbit Monoclonal Antibody [JB60-34]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human SET aa 160-290 / 290.
Species Reactivity
Human, Mouse
Validated Applications
WB, IF-Cell, IHC-P, IP, IF-Tissue
Molecular Weight
Predicted band size: 33 kDa
Positive Control
Rat kidney tissue, human tonsil tissue, mouse fallopian tube tissue, human lung carcinoma tissue, human kidney tissue, K-562 cell lysate, HeLa cell lysate, NIH/3T3 cell lysate.
Conjugation
unconjugated
Clone Number
JB60-34
RRID
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
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:500-1:2,000
-
IF-Cell
-
1:50-1:200
-
IHC-P
-
1:50-1:200
-
IP
-
1-2μg/sample
-
IF-Tissue
-
1:50
Target
Function
Multitasking protein, involved in apoptosis, transcription, nucleosome assembly and histone chaperoning. Isoform 2 anti-apoptotic activity is mediated by inhibition of the GZMA-activated DNase, NME1. In the course of cytotoxic T-lymphocyte (CTL)-induced apoptosis, GZMA cleaves SET, disrupting its binding to NME1 and releasing NME1 inhibition. Isoform 1 and isoform 2 are potent inhibitors of protein phosphatase 2A. Isoform 1 and isoform 2 inhibit EP300/CREBBP and PCAF-mediated acetylation of histones (HAT) and nucleosomes, most probably by masking the accessibility of lysines of histones to the acetylases. The predominant target for inhibition is histone H4. HAT inhibition leads to silencing of HAT-dependent transcription and prevents active demethylation of DNA. Both isoforms stimulate DNA replication of the adenovirus genome complexed with viral core proteins; however, isoform 2 specific activity is higher.
Background References
1. Beresford P J et al. Granzyme A activates an endoplasmic reticulum-associated caspase-independent nuclease to induce single-stranded DNA nicks. J Biol Chem 276:43285-43293 (2001).
2. Fan Z et al. Tumor suppressor NM23-H1 is a granzyme A-activated DNase during CTL-mediated apoptosis, and the nucleosome assembly protein SET is its inhibitor. Cell 112:659-672 (2003).
Sequence Similarity
Belongs to the nucleosome assembly protein (NAP) family.
Tissue Specificity
Widely expressed. Low levels in quiescent cells during serum starvation, contact inhibition or differentiation. Highly expressed in Wilms' tumor.
Post-translational Modification
Isoform 2 is phosphorylated on Ser-15 and Ser-24.; Isoform 2 is acetylated on Lys-11.; Some glutamate residues are glycylated by TTLL8. This modification occurs exclusively on glutamate residues and results in a glycine chain on the gamma-carboxyl group (By similarity).; N-terminus of isoform 1 is methylated by METTL11A/NTM1. Mainly trimethylated (By similarity).; [Isoform 2]: Cleaved after Lys-176 by GZMA. The cleavage inhibits its nucleosome assembly activity and disrupts the inhibition on NME1.
Subcellular Location
Nucleus. Cytoplasm. Endoplasmic reticulum.
Synonyms
2PP2A antibody
HLA DR associated protein II antibody
HLA-DR-associated protein II antibody
I 2PP2A antibody
I-2PP2A antibody
I2PP2A antibody
IGAAD antibody
Inhibitor of granzyme A activated DNase antibody
Inhibitor of granzyme A-activated DNase antibody
Inhibitor of GzmA-activated DNase antibody
ExpandImages
-
Western blot analysis of SET on different lysates with Rabbit anti-SET antibody (ET7107-18) at 1/1,000 dilution.
Lane 1: K-562 cell lysate
Lane 2: HeLa cell lysate
Lane 3: NIH/3T3 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 33 kDa
Observed band size: 40 kDa
Exposure time: 10 seconds; ECL: K1801;
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 (ET7107-18) 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 HeLa cells labeling SET with Rabbit anti-SET antibody (ET7107-18) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-SET antibody (ET7107-18) 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. -
Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-SET antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-18, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
Immunohistochemical analysis of paraffin-embedded mouse fallopian tube tissue using anti-SET antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-18, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-SET antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-18, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-SET antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-18, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-SET antibody (ET7107-18) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-18) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
SET was immunoprecipitated from 0.2 mg HeLa cell lysate with ET7107-18 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET7107-18 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: ET7107-18 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of ET7107-18 in HeLa cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"