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<strong>Figure 1 Western Blot Validation in Human 293 Cells</strong><br>Loading: 15 μg of lysates per lane.Antibodies: SAMHD1 (phospho Thr592) 8005 (1 μg/mL), (1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.293 cells were transfected with(A) empty expression vector,(B) wild-type SAMHD1,(C) SAMHD1 (mutation T592A) and(D) SAMHD1 (mutation T592E)
<strong>Figure 2 Mutant Validation in 293T Transfected Cells</strong><br>Loading: 15 μg of lysates per lane.Antibodies: SAMHD1 (phospho Thr592) 8005 (0.05 μg/mL), (1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.293 cells were transfected with (1) wild-type SAMHD1 or (2) SAMHD1 (mutation T592A).
<strong>Figure 3 Immunohistochemistry Validation of SAMHD1 (phospho Thr592) in Human Brain Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded Human Brain Tissue using anti- SAMHD1 (phospho Thr592) antibody (8005) at 2.5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4 °C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 4 Immunofluorescence Validation of SAMHD1 (phospho Thr592) in Human Brain Tissue</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed Human Brain Tissue labeling phospho SAMHD1 (phospho Thr592) with 8005 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
<strong>Figure 5 Mutant Validation of SAMHD1 (phospho Thr592) in human embryonic kidney (HEK) 293T cells (Antonucci et al., 2016) </strong><br>WB analysis showed protein expression level of SAMHD1 (phospho Thr592) detected by anti- SAMHD1 (phospho Thr592) antibody (8005). SAMHD1 (phospho Thr592) expression was detected only in SAMHD1 WT, but neither in SAMHD1 T592A mutant nor the control group (empty vector).
<strong>Figure 6 Mutant Validation of SAMHD1 (phospho Thr592) in primary monocyte-derived macrophages (MDM) from a healthy donor and human monocytic U937 cells (Antonucci et al., 2016) </strong><br>The expression levels of SAMHD1 (phospho Thr592) detected by anti- SAMHD1 (phospho Thr592) antibody (8005) in SAMHD1 mutants (D137N & Q548A) of U937 cells are lower than those of SAMHD1 WT or MDM.
<strong>Figure 7 Western Blot Validation of SAMHD1 (phospho Thr592) in herpes simplex viruses (HSV-1) infected human macrophages (Kueck et al., 2018) </strong><br>HSV-1 infected macrophages were treated with aryl hydrocarbon receptor (AhR) agonist or antagonist. WB analysis showed protein expression level of SAMHD1 (phospho Thr592) detected by anti- SAMHD1 (phospho Thr592) antibody (8005). AhR agonist repressed SAMHD1 (phospho Thr592) expression as compared to the control group or AhR antagonist. Representative of 2 different donors.
<strong>Figure 8 KD Validation of SAMHD1 (phospho Thr592) in human Macrophages (Mlcochova et al., 2017) </strong><br>SAMHD1 (phospho Thr592) signal detected by anti- SAMHD1 (phospho Thr592) antibody (8005) in SAMHD1 knockdown cells were disrupted as compared to control.
<strong>Figure 9 Western Blot Validation of SAMHD1 (phospho Thr592) in normal human fibroblasts (Tramentozzi et al., 2018) </strong><br>WB analysis showed protein expression level of SAMHD1 (phospho Thr592) detected by anti- SAMHD1 (phospho Thr592) antibody (8005). SAMHD1 (phospho Thr592) expression was detected in S and G2/M phase of cell cycle but not in G1 phase.
<strong>Figure 10 Regulated Expression Validation of SAMHD1 (phospho Thr592) in parental THP-1 cells (Tramentozzi et al., 2018) </strong><br>WB analysis showed protein expression level of SAMHD1 (phospho Thr592) detected by anti- SAMHD1 (phospho Thr592) antibody (8005). SAMHD1 (phospho Thr592) expression was significantly decreased in S and G2/M phase with the treatment of lambda-phosphatase.
<strong>Figure 11 Western Blot Validation of SAMHD1 (phospho Thr592) in Mouse SAMHD1 Transfected Cells (Wang et al., 2016) </strong><br>WB analysis showed protein expression level of SAMHD1 (phospho Thr592) detected by anti-SAMHD1 (phospho Thr592) antibody (8005). Mouse SAMHD1 was detected in transfected cells with WT mSAMHD1, but not with mutants.
<strong>Figure 12 Western Blot Validation of SAMHD1 (phospho Thr592) in Mouse Cells (Wang et al., 2016) </strong><br>WB analysis showed protein expression level of SAMHD1 (phospho Thr592) detected by anti-SAMHD1 (phospho Thr592) antibody (8005). Mouse SAMHD1 was detected mouse MEF cells
Figure 1 Western Blot Validation in Human 293 Cells
Loading: 15 μg of lysates per lane.Antibodies: SAMHD1 (phospho Thr592) 8005 (1 μg/mL), (1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.293 cells were transfected with(A) empty expression vector,(B) wild-type SAMHD1,(C) SAMHD1 (mutation T592A) and(D) SAMHD1 (mutation T592E)
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SAMHD1 (phospho Thr592) Antibody

CATALOG NUMBER: 8005

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Immunogen:
Anti-SAMHD1 (phospho Thr592) antibody (8005) was raised against a phosphorylated peptide corresponding to 18 amino acids near the carboxy terminus of human Phospho-SAMHD1 surrounding T592.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC, WB
Application Note:
WB: 0.05-1 μg/mL; IHC: 2.5 μg/mL; IF: 20 μg/mL.
Antibody validated: Western Blot in human and mouse samples, Immunohistochemistry and Immunofluorescence in human samples. All other applications and species not yet tested.
Specificity:
Phospho-SAMHD1 antibody will only detect the phosphorylated form of SAMHD1.
Positive Control 1:
Cat. No. 10-301 - Human Brain Tissue Slide
Predicted Molecular Weight:
Predicted: 72kD
Observed: 72 kD
Validation:

Mutant validation (Figure 2, 5, 6 & 7): Anti-SAMHD1 (phospho Thr592) antibody (8005) specificity was further verified by SAMHD1 (phospho Thr592) mutant. SAMHD1 (phospho Thr592) signal in 293T cells (figure 2) or HEK293T cells (figure 5 & 7) or U937 cells (figure 6) transfected with SAMHD1 (phospho Thr592) mutant was disrupted in comparison with that in cells transfected with the wild-type.

KD validation (Figure 8): SAMHD1 (phospho Thr592) expression detected by anit-SAMHD1 (phospho Thr592) antibodies (8005) was disrupted in knockdown macrophages.

Regulated expression validation (Figure 10): SAMHD1 (phospho Thr592) expression detected by anit-SAMHD1 (phospho Thr592) antibodies (8005) was down-regulated by lambda-phosphatase treatment in S and G2 phase of cell cycle.

Isoforms:
Human SAMHD1 (phospho Thr592) has 3 isoforms, including isoform 1 (626aa, 72kD), isoform 3 (556aa, 64kD), isoform 4 (591aa, 68kD). Mouse SAMHD1 has 2 isoforms, including (658aa, 76kD) and (651aa, 75kD). Rat SAMHD1 has only one isoform (620aa, 72kD). 8005 can detect human and mouse.
Purification:
SAMHD1 (phospho Thr592) antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
SAMHD1 (phospho Thr592) antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
SAMHD1 (phospho Thr592) antibody can be stored at 4°C for three months and -20°C, stable for up to one year.
Ncbi Official Symbol:
SAMHD1 (phospho Thr592)
Additional Names:
SAM domain and HD domain 1, DCIP, CHBL2, HDDC1, MOP-5, SBBI88
Protein Accession Number:
NP_056289
Protein Gi Number:
38016914
Ncbi Gene Id Number:
25939
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
The SAM domain and HD domain 1 (SAMHD1) protein is upregulated in response to viral infection and is thought to play a role in innate immunity (1). SAMHD1 blocks the infection of HIV-1 and SIVdeltaVpx before reverse transcription in macrophages and dendritic cells (2), and this restriction is regulated by phosphorylation of SAMHD1 (3). Mutations in this gene have been associated with Aicardi-Goutieres syndrome (1).
Background Reference 1:
Rice et al. Nat. Genet. 2009; 41:829-32.
Background Reference 2:
Hrecka et al. Nature 2011; 474:654-7.
Background Reference 3:
Welbourn et al. J. Virol. 2013; 87:11516-24.
Citation 1:
Antonucci et al. SAMHD1-mediated HIV-1 restriction in cells does not involvePMID: 27711056
Citation 2:
Kueck et al. The aryl hydrocarbon receptor and interferon gamma generate antiviral states via transcriptional repression. Elife. 2018;7. PMID: 30132758
Citation 3:
St Gelais et al. A Cyclin-Binding Motif in Human SAMHD1 Is Required for Its HIV-1 Restriction, dNTPase Activity, Tetramer Formation, and Efficient Phosphorylation. J Virol. 2018;92(6)PMID: 29321329
Citation 4:
Tramentozzi et al. The dNTP triphosphohydrolase activity of SAMHD1 persists during S-phase when the enzyme is phosphorylated at T592. Cell Cycle. 2018;17(9):1102-1114. PMID: 30039733
Citation 5:
Mlcochova et al. A G1-like state allows HIV-1 to bypass SAMHD1 restriction in macrophages. EMBO J. 2017;36(5):604-616.PMID: 28122869
Citation 6:
Mlcochova et al. DNA damage induced by topoisomerase inhibitors activates SAMHD1 and blocks HIV-1 infection of macrophages. EMBO J. 2018;37(1):50-62. PMID: 29084722
Citation 7:
Herrmann et al. The SAMHD1-mediated block of LINE-1 retroelements is regulated by phosphorylation. Mob DNA. 2018;9:11. PMID: 29610582
Citation 8:
Osei et al. USP18 (UBP43) Abrogates p21-Mediated Inhibition of HIV-1. J Virol. 2018;92(20). PMID: 30068654
Citation 9:
St Gelais et al. A Putative Cyclin-binding Motif in Human SAMHD1 Contributes to Protein Phosphorylation, Localization, and Stability. J Biol Chem. 2016;291(51):26332-26342. PMID: 27815502
Citation 10:
Wittmann et al. Phosphorylation of murine SAMHD1 regulates its antiretroviral activity. Retrovirology. 2015;12:103.PMID: 26667483
Citation 11:
Feifei Wang. Comparison of Murine and Human SAMHD1's Role in Retroviral Restriction and Cell Cycle Progression. PhD Thesis. The Ohio State University. 2015.
Citation 12:
Alexandra Herrmann. The role of SAMHD1 in restriction and immune sensing of retroviruses and retroelements. PhD Thesis. The Friedrich Alexander University Erlangen-Nuremberg. 2018.

FOR RESEARCH USE ONLY

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Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

8005

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

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