MADCAM1 anticorps (Middle Region)

N° du produit ABIN3044076

Cet anticorps anti-MADCAM1 Polyclonal Lapin (ABIN3044076) détecte spécifiquement MADCAM1 dans WB, IHC et IF. L’anticorps est réactif avec des échantillons de Humain. Validé indépendamment pour une utilisation en Immunohistochemistry.

Aperçu rapide pour MADCAM1 anticorps (Middle Region) (ABIN3044076)

Antigène: MADCAM1 (Mucosal Vascular Addressin Cell Adhesion Molecule 1 (MADCAM1))

Reactivité: Humain

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp MADCAM1 est non-conjugé

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

Détail du produit anti-MADCAM1 anticorps

Épitope: Middle Region

Fonction: Anti-MAdCAM1 Antibody

Séquence: EEPQGDEDVL FRVTERWR

Specificité: No cross reactivity with other proteins.

Réactivité croisée (Details): No cross-reactivity with other proteins

Attributs du produit: Anti-MAdCAM1 Antibody. Tested in IF, IHC, WB applications. This antibody reacts with Human.

Purification: Immunogen affinity purified.

Immunogène: A synthetic peptide corresponding to a sequence in the middle region of human MAdCAM1.

Isotype: IgG

Information d'application

Indications d'application: Western blot, 0.1-0.5 μg/mL, Human Immunohistochemistry (Paraffin-embedded Section), 2-5 μg/mL, Human Immunofluorescence, 5 μg/mL, Human

Commentaires:

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

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #300031 (Immunohistochemistry)

by: Human Protein Atlas

No.: #300031

Date: 13.12.2016

Antigène: MADCAM1

Application validée: Immunohistochemistry

Contrôle positif: colon, spleen

Conclusion: Passed. ABIN3044076 staining is consistent with RNA-seq data and published literature.

Validation Images

Staining of human colon (A.) and spleen (B.) sections in a tissue microarray with ABIN3044076. See the protocol for detailed information.

Protocole

Anticorps primaire: ABIN3044076

Anticorps secondaire: HRP polymer (ThermoFisher Scientific, TL-125-PH)

Full Protocol:

• Sample preparation:
• • Fixation and TMA preparation according to PMID: 22688270.
  • Cut 4µm TMA sections using a waterfall microtome (ThermoFisher Scientific, HM355S).
  • Dry sections ON at RT and then back them for 12-24h at 50°C.
• Deparaffinization and rehydration in xylene and graded alcohol series:
• • xylene twice for 3min.
  • 100% ethanol for 1min.
  • 95% ethanol for 1min.
  • 0.3% H₂O₂ in 95% ethanol for 5min to block endogenous peroxidase.
  • 70% ethanol for 1min.
  • 50% ethanol for 1min.
  • distilled H₂O for 2min.
• Antigen Retrieval by Heat Induced Epitope Retrieval (HIER):
• • Boil the sections in Lab Vision Citrate buffer pH6.0 (ThermoFisher Scientific, TA-250-PM1X) for 4min at 125°C in a decloaking chamber (Biocare Medical).
  • Allow slides to cool down to 90°C in the decloaking chamber.
• Immunostaining in a Lab Vision Autostainer 480 (ThermorFisher Scientific) at RT with 300µl of reagents for each step:
• • Rinse sections in Lab Vision wash buffer with extra tween added to a final concentration of 0.2% (ThermoFisher Scientific, TA-999-TT and TA-125-TW) wash buffer.
  • Block sections with Lab Vision Ultra V-Block (ThermoFisher Scientific, TA-125-UB) for 5min.
  • Rinse sections 2x in wash buffer.
  • Incubate sections with primary rabbit anti-MADCAM1 antibody (antibodies-online, ABIN3044076) diluted 1:900 for 30min.
  • Rinse sections 3x in wash buffer.
  • Incubate sections with labeled HRP polymer (ThermoFisher Scientific, TL-125-PH) for 30min.
  • Rinse sections 2x in wash buffer.
  • Develop in DAB Quanto solution (ThermoFisher Scientific, TL-125-QHDX) for 5min.
  • Rinse sections in distilled H₂O.
• Washing, counterstaining, and coverslipping in Autostainer XL (Leica Biosystems, ST5010) at RT:
• • Counterstaining in Mayer’s hematoxylin plus (HistoLab, 01820) for 7.5min.
  • Rinse sections in tap water for 5min.
  • Rinse sections in lithium carbonate water diluted 1:5 for 5min.
  • Dehydration in graded ethanol series and Neo-Clear (Merck Millipore, 109843) according to the manufacturer recommendations.
  • Automated mounting (Leica Biosystems, CV5030) of coverslip with Pertex (Histolab, 00871).
• Microscopy:
• • Image acquisition on a Leica Aperio AT2 and Aperio Scanscope AT at 20x magnification.

Notes: ABIN3044076 shows nice staining in a subset of vessels (endothelial cells), present in intestine and spleen.

Stockage

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 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.

Conseil sur la manipulation: Avoid repeated freezing and thawing.

Stock: 4 °C,-20 °C

Stockage commentaire: 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.

Date de péremption: 12 months

Détails sur MADCAM1

Antigène: MADCAM1 (Mucosal Vascular Addressin Cell Adhesion Molecule 1 (MADCAM1))

Autre désignation: MADCAM1

Sujet:

Background: MADCAM1 (Mucosal Vascular Addressin Cell Adhesion Molecule 1), also known as MACAM1, is a protein that in humans is encoded by the MADCAM1 gene. By PCR-based analysis of somatic cell hybrids, Leung et al. (1997) mapped the MACAM1 gene to chromosome 19. The protein encoded by this gene is an endothelil cell adhesion molecule that interacts preferentially with the leukocyte beta7 integrin LPAM-1 (alpha4 / beta7), L-selectin, and VLA-4 (alpha4/beta1) on myeloid cells to leukocytes into mucosal and inflamed tissues. It is a member of the immunoglobulin superfamily and is similar to ICAM-1 and VCAM-1.

Gene Full Name: mucosal vascular addressin cell adhesion molecule 1

Poids moléculaire: 60 kDa

ID gène: 8174

UniProt: Q13477