IdU anticorps

Détails pour le produit réf. ABIN2669973
Antigène
Reactivité
Chemical
8
5
Hôte
Souris
8
5
Clonalité (Clone)
Monoclonal ()
Application
Immunofluorescence (IF), Immunohistochemistry (IHC)
13
5
Options
'Independent Validation' signe
Antigène IdU
Numéro du lot W001
Application validée Immunohistochemistry
Contrôle positif

Adult (24 months) mouse brain and intestine (duodenum)

Contrôle négative

We incubated slices overnight with the blocking solution only and then processed them with the secondary antibody.

Conclusion

Passed. ABIN2669973 works in IHC-P with a microwave antigen retrieval pretreatment and concentration of 1:50.

Anticorps primaire ABIN2669973
Anticorps secondaire goat anti-mouse AF594-conjugated (Invitrogen by Thermo Fisher Scientific, A11032, lot 819561)
Protocole
  • Inject mouse intraperitoneally at the dose of 0.057mg/g body weight with IdU (Sigma, I7125) dissolved in sterile distilled water at final concentration of 100mg/ml, pH7.5.
  • Perfuse mouse with 4% paraformaldehyde in 0.1M phosphate buffer (PB) pH7.4 and post-fix samples in the same fixative for an additional 2h at RT.
  • Wash, dehydrate, and embed samples in paraffin wax.
  • Following several washes in PBS, cut intestines and brain with a microtome (6µm-thick sections) and mount sections on glass slides.
  • After paraffin removal, process sections by microwave antigen retrieval for 10min (95-100°C) in 10mM sodium citrate buffer pH6.0.
  • Let sections cool for 20min.
  • Wash sections 2x 5min with distilled water.
  • Wash sections 5min with PBS.
  • Incubate sections in PBS containing 1% albumin from chicken egg white (Sigma, A5378) and 0.3% Triton-X-100 (BioRad, 161-0407, lot 00583) for 1h at RT to block non-specific binding sites.
  • Incubate sections with primary antibody mouse anti-IdU (antibodies-online, ABIN2669973, lot W001) diluted 1:50 in PBS-BSA-PLL ON at 4°C.
  • Wash sections 3x 5min with 0.01M PBS.
  • Incubate sections with secondary goat anti-mouse AF594-conjugated (Invitrogen by Thermo Fisher Scientific, A11032, lot 819561) diluted 1:500 in 0.1M PBS for 1h at RT.
  • Wash sections 3x 5min with 0.01M PBS.
  • Mount sections in Fluoroshield (Sigma, F6182, lot MKCB0153V).
  • Acquire images with Leica DM 6000B fluorescence microscope equipped with the manufacturer’s FITC filter set and digital camera at magnifications of 20x and 40x. Parameters for image acquisition (Exposure 2.2s, Gain 2.3x, Saturation 1.5, Gamma 1.10) were maintained unchanged for all images.
Notes

For the incubation with the primary antibody ABIN2669973 and 1:20, 1:70, and 1:100 dilutions were also tested with 0.01M PBS, 5% Normal Goat Serum (NGS Sigma, G9023, lot SLBV1396) and 0.1% Triton-X -100 as the blocking solution, but 1:50 and PBS containing 1% albumin from chicken egg white and 0.3% Triton-X gave the best results.

Validation Images
Immunohistochemistry validation image for anti-IdU antibody (ABIN2669973) Staining of mouse brain and intestine sections using ABIN2669973. A-B: A bright stain...
Immunogène Iododeoxyuridine coupled to keyhole limpet hemocyanin.
Clone OTI2B10
Isotype IgG2b
Purification Purified from mouse ascites fluids by affinity chromatography
Antigène
Indications d'application IHC 1:150, IF 1:150,
Restrictions For Research Use only
Format Liquid
Concentration 4.54 mg/mL
Images (Fournisseur)
Immunofluorescence (IF) image for anti-IdU antibody (ABIN2669973) anti-IdU antibody
Immunohistochemistry (IHC) image for anti-IdU antibody (ABIN2669973) anti-IdU antibody (Image 2)
Immunohistochemistry (IHC) image for anti-IdU antibody (ABIN2669973) anti-IdU antibody (Image 3)
Immunohistochemistry (IHC) image for anti-IdU antibody (ABIN2669973) anti-IdU antibody (Image 4)
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