SAMHD1 Recombinant Rabbit Monoclonal Antibody [JU56-04]
Catalog# ET1706-24
SAMHD1 Recombinant Rabbit Monoclonal Antibody [JU56-04]
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WB
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IF-Cell
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IHC-P
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FC
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Human
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unconjugated
Overview
Product Name
SAMHD1 Recombinant Rabbit Monoclonal Antibody [JU56-04]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human SAMHD1 aa 577-626 / 626.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IHC-P, FC
Molecular Weight
72 kDa
Positive Control
THP-1 cell lysates, HepG2, HUVEC, LOVO, human tonsil tissue tissue, human colon carcinoma tissue, human spleen tissue, K562.
Conjugation
unconjugated
Clone Number
JU56-04
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
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WB
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1:1,000-1:2,000
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IF-Cell
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1:100-1:500
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IHC-P
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1:50-1:200
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FC
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1:50-1:100
Target
Function
SAM domain and HD domain-containing protein 1 is a protein that in humans is encoded by the SAMHD1 gene. SAMHD1 is a cellular enzyme, responsible for blocking replication of HIV in dendritic cells, macrophages and monocytes. It is an enzyme that exhibits phosphohydrolase activity, converting deoxynucleoside triphosphates (dNTPs) to inorganic phosphate (iPPP) and a 2'-deoxynucleoside (i.e. deoxynucleosides without a phosphate group). In doing so, SAMHD1 depletes the pool of dNTPs available to a reverse transcriptase for viral cDNA synthesis and thus prevents viral replication. SAMHD1 has also shown nuclease activity. Although a ribonuclease activity was described to be required for HIV-1 restriction, recent data confirmed that SAMHD1-mediated HIV-1 restriction in cells does not involve ribonuclease activity.
Background References
1. Mlcochova P et al. A G1-like state allows HIV-1 to bypass SAMHD1 restriction in macrophages. EMBO J 36:604-616 (2017).
2. Badia R et al. The G1/S Specific Cyclin D2 Is a Regulator of HIV-1 Restriction in Non-proliferating Cells. PLoS Pathog 12:e1005829 (2016).
Sequence Similarity
Belongs to the SAMHD1 family.
Tissue Specificity
Expressed in heart, skeletal muscle, spleen, liver, small intestine, placenta, lung and peripheral blood leukocytes. No expression is seen in brain and thymus.
Post-translational Modification
Phosphorylation at Thr-592 by CDK1 acts as a switch to control deoxynucleoside triphosphate (dNTPase)-dependent and -independent functions. Phosphorylation at Thr-592 takes place in cycling cells: it reduces the stability of the homotetramer, impairing the dNTPase activity and subsequent ability to restrict infection by viruses. It also inhibits ability to suppress LINE-1 retrotransposon activity. In contrast, phosphorylation at Thr-592 promotes DNA end resection at stalled replication forks in response to DNA damage.; (Microbial infection) Ubiquitinated following interaction with HIV-2 viral protein Vpx; Vpx promotes interaction and with a DCX (DDB1-CUL4-X-box) E3 ubiquitin ligase, leading to proteasomal degradation.
Subcellular Location
Nucleus.
UNIPROT
Synonyms
CHBL2 antibody
DCIP antibody
Dendritic cell derived IFNG induced protein antibody
Dendritic cell-derived IFNG-induced protein antibody
Deoxynucleoside triphosphate triphosphohydrolase SAMHD1 antibody
dNTPase antibody
HD domain containing 1 antibody
HDDC1 antibody
Mg11 antibody
Monocyte protein 5 antibody
ExpandImages
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Western blot analysis of SAMHD1 on THP-1 cell lysates with Rabbit anti-SAMHD1 antibody (ET1706-24) at 1/500 dilution.
Lysates/proteins at 10 µg/Lane.
Predicted band size: 72 kDa
Observed band size: 72 kDa
Exposure time: 30 seconds;
8% 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 (ET1706-24) 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. -
Immunocytochemistry analysis of HepG2 cells labeling SAMHD1 with Rabbit anti-SAMHD1 antibody (ET1706-24) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-SAMHD1 antibody (ET1706-24) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. -
Immunocytochemistry analysis of HUVEC cells labeling SAMHD1 with Rabbit anti-SAMHD1 antibody (ET1706-24) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-SAMHD1 antibody (ET1706-24) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. -
Immunocytochemistry analysis of LOVO cells labeling SAMHD1 with Rabbit anti-SAMHD1 antibody (ET1706-24) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-SAMHD1 antibody (ET1706-24) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. -
Immunohistochemical analysis of paraffin-embedded human tonsil tissue tissue with Rabbit anti-SAMHD1 antibody (ET1706-24) at 1/50 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1706-24) at 1/50 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. -
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-SAMHD1 antibody (ET1706-24) at 1/50 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1706-24) at 1/50 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. -
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-SAMHD1 antibody (ET1706-24) at 1/50 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1706-24) at 1/50 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. -
Flow cytometric analysis of SAMHD1 was done on K562 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1706-24, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"