- PA5-23381 - Invitrogen Antibodies STING1 antibody

Submitted references Human rhinovirus promotes STING trafficking to replication organelles to promote viral replication.

Triantafilou M, Ramanjulu J, Booty LM, Jimenez-Duran G, Keles H, Saunders K, Nevins N, Koppe E, Modis LK, Pesiridis GS, Bertin J, Triantafilou K
Nature communications 2022 Mar 17;13(1):1406

ASA404, a vascular disrupting agent, as an experimental treatment approach for brain tumors.

Bähr O, Gross S, Harter PN, Kirches E, Mawrin C, Steinbach JP, Mittelbronn M
Oncology letters 2017 Nov;14(5):5443-5451

AIM2-Like Receptors Positively and Negatively Regulate the Interferon Response Induced by Cytosolic DNA.

Nakaya Y, Lilue J, Stavrou S, Moran EA, Ross SR
mBio 2017 Jul 5;8(4)

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-TMEM173 (Product # PA5-23381 ) and a 38 kDa band corresponding to TMEM173 was observed. Whole cell extracts (30 µg lysate) of THP-1 (Lane 1), U-937 (Lane 2), HeLa (Lane 3), Jurkat (Lane 4) and K-562 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein 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 (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20,000 using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of STING in THP-1, HT-29, U2OS cells and human spleen. Samples were incubated in STING polyclonal antibody (Product # PA5-23381) using a dilution of 2 µg/mL followed by an anti-rabbit HRP secondary antibody. Separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. Detection: chemiluminescence.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of TMEM173 was achieved by transfecting THP-1 with TMEM173 specific siRNAs (Silencer® select Product # S50646, S50645). Western blot analysis (Fig. a) was performed using Whole cell extracts from the TMEM173 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with TMEM173 (Product # PA5-23381 , 1:3000 ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to TMEM173.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry analysis of STING in RH-30 cells fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton X-100. Samples were incubated in STING polyclonal antibody (Product # PA5-23381) using a dilution of 2 µg/mL overnight at 4 °C followed by anti-rabbit DyLight 488 (Green) with a dilution of 1:500. Alpha Tubulin Antibody (DM1A) was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of STING in HT-29 cells fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. Samples were incubated in STING polyclonal antibody (Product # PA5-23381) using a dilution of 10 µg/mL overnight at 4 °C followed by anti-rabbit DyLight 488 (Green) with a dilution of 1:500. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of TMEM173 was performed using 70% confluent log phase U-2 OS cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.01% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with TMEM173 (Product # PA5-23381 ) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image predominantly showing endoplasmic reticulum localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7C1115LZR). magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry of STING in RH30 cells (blue) and a matched isotype control (orange). Samples were incubated in STING polyclonal antibody (Product # PA5-23381) using a dilution of 2.5 µg/mL for 30 minutes at room temperature followed by a Rabbit IgG (H+L) Cross-Adsorbed secondary antibody. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry of STING in THP-1 cells and a matched isotype control. Samples were incubated in STING polyclonal antibody (Product # PA5-23381) using a dilution of 2.5 µg/mL for 30 minutes at room temperature followed by a Rabbit IgG (H+L) Cross-Adsorbed secondary antibody. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry of STING in U937 cells. Samples were incubated in STING polyclonal antibody (Product # PA5-23381) using a dilution of 2.5 µg/mL for 30 minutes at room temperature. Antibody (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Both antibodies were conjugated to Alexa Fluor 594.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIG 7 IFI205-STING interaction increases upon TREX1 and AIM2 depletion. (A) PLA for interactions of IFI205myc-AIM2, IFI205myc-STING, and AIM2-STING in NR9456-IFI205myc or NR9456 cells. The number of cells and images quantified for each condition in NR9456-IFI205myc cells are as follows: IFI205myc-Sting, 141 cells and 10 images; IFI205myc-Aim2, 101 cells and 6 images; control, 65 cells and 5 images. The number of cells and images quantified for each condition in NR9456 cells are as follows: control, 74 cells and 5 images; Aim2-Sting, 47 cells and 5 images. Controls for knockdowns are shown in Fig. S9A . (B) PLA for IFI205myc-STING interactions in NR9456-IFI205myc cells with knockdown of the genes indicated. Quantification: siCont, 102 cells and 5 images; siTrex1, 190 cells and 7 images; siTrex1+siAim2, 108 cells and 5 images. A representative image is shown. This experiment was repeated twice and gave similar results both times. (C) PLA for AIM2-STING interaction in NR9456 cells after knockdown of the genes indicated. Quantification: siCont, 84 cells and 4 images; siTrex1, 33 cells and 4 images; siTrex1+si205, 58 cells and 4 images. PLA dots were quantified and normalized to cell numbers based on DAPI staining with ImageJ software. The values shown are the mean +- the standard error of the mean of different pictures. **, P < 0.005; ***, P < 0.0005. (two-tailed t test). siCont, control siRNA.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 8. STING expression and iba1 positive cells in subcutaneous and intracranial U-87 tumors. Immunohistochemistry for STING is shown for subcutaneous (A) and intracranial (B) U-87 tumors. Cells positive for iba1 are stained in subcutaneous (C) and intracranial (D) U-87 tumors.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 5 STIM1 releases STING upon HRV infection. FRET studies measuring donor (STING) and acceptor (ERGIC or STIM1) interactions in BEAS-2B cells infected with HRV for 2 h or exposed to ISD (1 ug) ( a ). Confocal microscopy of BEAS-2B cells infected with HRV for 2 h assessing colocalization between STIM1, the ER and STING. Data in a is means +/- SD ( n = 3, independent experiments). Statistical significance between unstimulated and virus-infected conditions was assessed by unpaired student's t -test. *** p < 0.001 Data in b is representative of three biological donors with at least 20 technical replicates. Degree of colocalization is presented at R(obs). Bars shown at 10 um.

PA5-23381

Antibody data