Invitrogen

SAMHD1 Monoclonal Antibody (OTI1A1)

Advanced Verification

This Antibody was verified by Knockout to ensure that the antibody binds to the antigen stated.

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Datasheet

Protocols

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SAMHD1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of SAMHD1 was performed using 70% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with SAMHD1 Monoclonal Antibody (OTI1A1) (Product # MA5-25354) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear and plasma membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

SAMHD1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry was performed on paraffin-embedded adenocarcinoma of human ovary tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a SAMHD1 monoclonal antibody (Product # MA5-25354).

SAMHD1 Antibody in Western Blot (WB) — Western blot was performed using Anti-SAMHD1 Monoclonal Antibody (OTI1A1) (Product # MA5-25354) and a 72 kDa band corresponding to SAMHD1 was observed in all cell and tissue lysates tested except Jurkat and U-937. Lower expression of SAMHD1 was observed in Mouse Brain as compared to Mouse Lung and Mouse Spleen [10.1016/s0165-2478(00)00276-5]. Modified whole cell extracts (1% SDS) (30 µg lysate) of THP-1 (Lane 1), Caco-2 (Lane 2), Jurkat (Lane 3), U-937 (Lane 4), Mouse Lung (Lane 5), Mouse Spleen (Lane 6), Rat Spleen (Lane 7), Mouse Brain (Lane 8) and Rat Brain (Lane 9) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:200 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A ~25 kDa band corresponding to tissue IgG was observed in some mouse tissue models.

SAMHD1 Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. Loss of signal was observed for target protein in SAMHD1 knockout (KO) cell line using Anti-SAMHD1 polyclonal antibody (Product # MA5-25354). {KO}

Product Details

MA5-25354

Applications	Tested Dilution	Publications
Western Blot (WB)	1:200-1:2,000	-
Immunohistochemistry (Paraffin) (IHC (P))	1:150	-
Immunocytochemistry (ICC/IF)	1:100	-
Flow Cytometry (Flow)	1:100	-

Product Specifications
Species Reactivity	Dog, Human, Mouse, Non-human primate, Rat
Host/Isotype	Mouse / IgG2b
Class	Monoclonal
Type	Antibody
Clone	OTI1A1
Immunogen	Full length human recombinant protein of SAMHD1 produced in HEK293T cell View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1.0 mg/mL
Purification	Affinity Chromatography
Storage buffer	PBS, pH 7.3, with 1% BSA, 50% glycerol
Contains	0.02% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2725288

Target Information

This gene may play a role in regulation of the innate immune response. The encoded protein is upregulated in response to viral infection and may be involved in mediation of tumor necrosis factor-alpha proinflammatory responses. Mutations in this gene have been associated with Aicardi-Goutieres syndrome.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: DCIP; Dendritic cell-derived IFNG-induced protein; Deoxynucleoside triphosphate triphosphohydrolase SAMHD1; dNTPase; hSAMHD1; IFN-gamma induced; Interferon-gamma-inducible protein Mg11; Monocyte protein 5; MOP-5; mSAMHD1; RP1-132F21.1; SAM domain and HD domain 1; SAM domain and HD domain-containing protein 1

Gene Aliases: CHBL2; DCIP; E330031J07Rik; HDDC1; Mg11; Mg21; MOP-5; MOP5; SAMHD1; SBBI88

UniProt ID: (Human) Q9Y3Z3, (Mouse) Q60710

Entrez Gene ID: (Human) 25939, (Dog) 485862, (Rat) 311580, (Mouse) 56045

Molecular Function: hydrolase