MADCAM1 Antikörper (Middle Region)

Produktnummer ABIN3044076

Der Kaninchen Polyklonal anti-MADCAM1 Antikörper (ABIN3044076) detektiert spezifisch MADCAM1 in WB, IHC und IF. Dieser Antikörper reagiert spezifisch mit Proben aus Human. Er hat eine unabhängige Validierung für Immunohistochemistry.

Kurzübersicht für MADCAM1 Antikörper (Middle Region) (ABIN3044076)

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

Reaktivität: Human

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser MADCAM1 Antikörper ist unkonjugiert

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

Produktdetails anti-MADCAM1 Antikörper

Bindungsspezifität: AA 173-190, Middle Region

Verwendungszweck: Anti-MAdCAM1 Antibody

Sequenz: EEPQGDEDVL FRVTERWR

Kreuzreaktivität (Details): No cross-reactivity with other proteins

Produktmerkmale: Anti-MAdCAM1 Antibody (ABIN3044076). Tested in IF, IHC, WB applications. This antibody reacts with Human. This is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications.

Aufreinigung: Immunogen affinity purified.

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

Isotyp: IgG

Anwendungsinformationen

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

Kommentare:

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

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #300031 (Immunohistochemistry)

by: Human Protein Atlas

No.: #300031

Datum: 13.12.2016

Antigen: MADCAM1

Validierte Anwendung: Immunohistochemistry

Positivkontrolle: colon, spleen

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

Validierungsbilder

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

Protokoll

Primärantikörper: ABIN3044076

Sekundärantikörper: 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.

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

Handhabung

Format: Lyophilized

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

Konzentration: 500 μg/mL

Buffer: Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.

Handhabung: Avoid repeated freezing and thawing.

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: 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.

Haltbarkeit: 12 months

Details zu MADCAM1

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

Andere Bezeichnung: MADCAM1

Hintergrund:

Synonyms: Mucosal addressin cell adhesion molecule 1,MAdCAM-1,hMAdCAM-1,MADCAM1,

Tissue Specificity: Highly expressed on high endothelial venules (HEV) and lamina propia venules found in the small intestine, and to a lesser extent in the colon and spleen. Very low levels of expression found in pancreas and brain. Not expressed in the thymus, prostate, ovaries, testis, heart, placenta, lung, liver, skeletal muscle, kidney or peripheral blood leukocytes. .

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.

Molekulargewicht: 60 kDa

UniProt: Q13477