STING/TMEM173 antibody (C-Term)

Catalog No. ABIN3043423

This independently validated Rabbit Polyclonal antibody specifically detects STING/TMEM173 in WB and IHC. It exhibits reactivity toward Human.

Quick Overview for STING/TMEM173 antibody (C-Term) (ABIN3043423)

Target: STING/TMEM173 (TMEM173) (Transmembrane Protein 173 (TMEM173))

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This STING/TMEM173 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC)

Product Details anti-STING/TMEM173 Antibody

Binding Specificity: AA 284-316, C-Term

Purpose: Anti-TMEM173 Antibody Picoband®

Sequence: RLEQAKLFCR TLEDILADAP ESQNNCRLIA YQE

Cross-Reactivity (Details): No cross-reactivity with other proteins

Characteristics: Anti-TM Antibody Picoband® (ABIN3043423). Tested in IHC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.

Purification: Immunogen affinity purified.

Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human TMEM173, different from the related mouse sequence by five amino acids.

Isotype: IgG

Application Details

Application Notes: Immunohistochemistry (Paraffin-embedded Section), 0.5-1 μg/mL, Human
Western blot, 0.1-0.5 μg/mL, Human
1. "Entrez Gene: Transmembrane protein 173". 2. Ishikawa, H, Barber, G. N. (2008). "STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling".Nature 455 (7213): 674-8. 3. Nazmi, A, Mukhopadhyay, R, Dutta, K, Basu, A (2012)."STING mediates neuronal innate immune response following Japanese encephalitis virus infection". Scientific Reports 2: 347.

Comment:

Antibody can be supported by chemiluminescence kit ABIN921124 in WB, supported by ABIN921231 in IHC(P).

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #100035 (Immunohistochemistry)

by: University of California, Los Angeles

No.: #100035

Date: 07/02/2016

Antigen: Anti-TMEM173 Picoband™ Antibody

Lot Number: 0951512Da071365

Method validated: Immunohistochemistry

Positive Control: Human pancreatic adenocarcinoma (PDAC)

Notes: In primary human pancreatic tumor tissue, ABIN3043423 stains specifically the tumor cell cytoplasm only, not fibroblasts.

Validation Images

Immunohistochemistry on pancreatic adenocarcinoma (PDAC) FFPE tissue sections. The primary TMEM173 antibody ABIN3043423 was used 1:500 diluted in 5% normal Goat serum in PBS with a biotin-donkey-anti-rabbit IgG (H+L) secondary antibody. A. IHC staining of PDAC tissue from patient #1 at 400x magnification. B. IHC staining of PDAC tissue from patient #2 at 200x magnification.

Full Methods

Primary Antibody: ABIN3043423

Secondary Antibody: Biotin-SP-AffinPure Donkey-anti-rabbit IgG (H+L), Jackson ImmunoResearch, cat#711-065-152

Full Protocol:

• Deparaffinize slides
• • Bake slides in oven at 60°C for 1h and let cool completely to RT.
  • Rehydrate:
  • Xylene 3x 5min
  • 100% EtOH 2x 2min
  • 95% EtOH 3x 2min
  • 70% EtOH 1x 2min
  • 50% EtOH 1x 2min
  • H2O 2x 3min
• Blocking peroxidase activity
• • Treat in 3% H2O2-PBS for 15min on rotator at RT (240 ml/slide hold chamber).
  • Wash with PBS 3x 2min.
• Antigen retrieval
• • Citrate buffer stock solution 100x, pH6.0 working solution 0.01M, freshly diluted into working solution.
  • Boil Citrate buffer until 100°C on hot plate, put slides in the boiled buffer, keep boiling 15min, then let them cool down on bench top for 20min.
  • Wash with H2O 2x 2min.
  • Wash with PBS 3x 5min.
  • PAP-pen cycles the slides: using vacuum to suck off the solution by cycling around the tissue area, then using the PAP-pen draw along the cycle line. Make sure the tissue area is kept wet.
• Apply blocking solution
• • Incubate with 50-100µl (cover the whole tissue area) 5% donkey serum in PBS for 1h in moist a box at RT.
  • Blocking stock solution: 5% donkey serum in 10 ml PBS.
  • Drain blocking solution and blot excess liquid with Kim wipe.
  • Prepare primary antibody solution in blocking buffer.
• Apply primary antibody
• • Dilute primary TMEM173 antibody ABIN3043423 1:500 dilution in 5% normal goat serum in PBS.
  • Incubate overnight in a box at 4°C to assure amoist environment and prevent slides from drying.
• Wash with 0.05% Tween-PBS3x 5min.
• Dilute Biotin-SP-AffinPure Donkey-anti-rabbit IgG (H+L) secondary antibody with 5% blocking solution (5% donkey serum)
• Incubate 1h with secondary antibody at room temperature.
• Prepare ABC solution 1:200: dilute both A and B in 0.05% Tween-PBS. Allow ABC diluted solution to sit for 30-60min before using, keep in the dark.
• Wash with 0.05% Tween-PBS 5min, 7min, and 7min.
• Apply 250µL/slide ABC solution; incubate for 30min at RT in moist incubation box.
• Wash with 0.05% Tween-PBS 5min, 7min, and 7min.
• Filter Hematoxylin.
• Prepare fresh DAB solution in disposable beaker (do not allow solution to sit):
• • 2.5ml H2O + 1 drop buffer + 2 drops DAB + 1 drop H2O2
  • Use transfer pipette to apply DAB x 1min
• Wash 3x with dH2O (10 dips each).
• Hematoxylin stain 5-10sec.
• Wash until water is clear.
• Hematoxylin stain 5-10sec.
• Dehydrate
• • 50% EtOH 1x 2min.
  • 70% EtOH 1x 2min.
  • 95% EtOH 2x 2min.
  • 100% EtOH 2x 2min.
  • Xylene 3x 5min.
• Apply cover-slip. Allow glue to dry overnight.

Handling

Format: Lyophilized

Reconstitution: Add 0.2 mL of distilled water will yield a concentration of 500 μg/mL.

Concentration: 500 μg/mL

Buffer: Each vial contains 5 mg BSA, 0.9 mg NaCl, 0.2 mg Na2HPO4, 0.05 mg Sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freezing and thawing.

Storage: 4 °C,-20 °C

Storage Comment: Store at -20°C for one year from date of receipt. After reconstitution, at 4°C for one month.
It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freeze-thaw cycles.

Target Details for STING/TMEM173

Target: STING/TMEM173 (TMEM173) (Transmembrane Protein 173 (TMEM173))

Alternative Name: TMEM173

Background:

Synonyms: Stimulator of interferon genes protein,hSTING,Endoplasmic reticulum interferon stimulator,ERIS,Mediator of IRF3 activation,hMITA,Transmembrane protein 173,TMEM173,ERIS, MITA, STING,

Tissue Specificity: Ubiquitously expressed. Expressed in skin endothelial cells, alveolar type 2 pneumocytes, bronchial epithelium and alveolar macrophages. .

Background: Transmembrane protein 173 is a protein that in humans is encoded by the TMEM173 gene. This gene encodes a five transmembrane protein that functions as a major regulator of the innate immune response to viral and bacterial infections. The encoded protein is a pattern recognition receptor that detects cytosolic nucleic acids and transmits signals that activate type I interferon responses. Also the encoded protein has been shown to play a role in apoptotic signaling by associating with type II major histocompatibility complex. Mutations in this gene are the cause of infantile-onset STING-associated vasculopathy. Alternate splicing results in multiple transcript variants.

Sequence Similarities: Belongs to the transcription factor STAT family.

Molecular Weight: 42 kDa

Gene ID: 340061

Pathways: Activation of Innate immune Response