Mucosal Vascular Addressin Cell Adhesion Molecule 1 (MADCAM1) (AA 173-190), (Middle Region) antibody

Details for Product No. ABIN3044076, Supplier: Log in to see
Antigen
  • MADCAM1
  • AV211525
  • MACAM1
  • mucosal vascular addressin cell adhesion molecule 1
  • MADCAM1
  • Madcam1
  • MADCAM-1
Epitope
AA 173-190, Middle Region
20
5
3
3
1
1
1
1
1
1
1
Reactivity
Human
131
100
33
2
Host
Rabbit
108
61
44
1
Clonality
Polyclonal
Conjugate
Un-conjugated
25
12
9
7
5
5
5
5
5
4
3
2
2
2
2
2
2
2
2
2
2
2
2
1
1
1
1
Application
Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
115
113
45
43
35
29
26
22
10
8
7
3
3
2
2
2
2
2
2
1
1
1
1
Options
Supplier
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'Independent Validation' Badge
Antigen MADCAM1
Method validated Immunohistochemistry
Positive Control colon, spleen
Notes Passed. ABIN3044076 staining is consistent with RNA-seq data and published literature.
Primary Antibody ABIN3044076
Secondary Antibody HRP polymer (ThermoFisher Scientific, TL-125-PH)
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.
Experimental Notes ABIN3044076 shows nice staining in a subset of vessels (endothelial cells), present in intestine and spleen.
Validation Images
Immunohistochemistry validation image for anti-Mucosal Vascular Addressin Cell Adhesion Molecule 1 (MADCAM1) (AA 173-190), (Middle Region) antibody (ABIN3044076) Staining of human colon (A.) and spleen (B.) sections in a tissue microarray with ABI...
Purpose Rabbit IgG polyclonal antibody for Mucosal addressin cell adhesion molecule 1(MADCAM1) detection. Tested with WB, IHC-P, IHC-F in Human.
Immunogen A synthetic peptide corresponding to a sequence in the middle region of human MAdCAM1(173-190aa EEPQGDEDVLFRVTERWR).
Sequence EEPQGDEDVL FRVTERWR
Isotype IgG
Cross-Reactivity (Details) No cross reactivity with other proteins.
Characteristics Rabbit IgG polyclonal antibody for Mucosal addressin cell adhesion molecule 1(MADCAM1) detection. Tested with WB, IHC-P, IHC-F in Human.
Gene Name: mucosal vascular addressin cell adhesion molecule 1
Protein Name: Mucosal addressin cell adhesion molecule 1
Purification Immunogen affinity purified.
Alternative Name MADCAM1 (MADCAM1 Antibody Abstract)
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 direct leukocytes into mucosal and inflamed tissues. It is a member of the immunoglobulin superfamily and is similar to ICAM-1 and VCAM-1.

Synonyms: Addressin mucosal antibody|hMAdCAM 1 antibody|hMAdCAM-1 antibody|MACAM1 antibody|MADCA_HUMAN antibody|MAdCAM 1 antibody|MAdCAM-1 antibody|Madcam1 antibody|Mucosal addressin cell adhesion molecule 1 antibody|Mucosal vascular addressin cell adhesion molecule 1 antibody
UniProt Q13477
Application Notes WB: Concentration: 0.1-0.5 μg/mL, Tested Species: Human, The detection limit for MADCAM1 is approximately 0.5 ng/lane under reducing conditions.
IHC-P: Concentration: 0.5-1 μg/mL, Tested Species: Human, Epitope Retrieval by Heat: Boiling the paraffin sections in 10 mM citrate buffer, pH 6.0, for 20 mins is required for the staining of formalin/paraffin sections.
IHC-F: Concentration: 0.5-1 μg/mL, Tested Species: Human
Notes: Tested Species: Species with positive results. Predicted Species: Species predicted to be fit for the product based on sequence similarities. Other applications have not been tested. Optimal dilutions should be determined by end users.
Comment

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

Restrictions For Research Use only
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 Thimerosal, 0.05 mg Sodium azide.
Preservative Thimerosal (Merthiolate), Sodium azide
Precaution of Use This product contains Sodium azide and Thimerosal (Merthiolate): POISONOUS AND HAZARDOUS SUBSTANCES which should be handled by trained staff only.
Handling Advice Avoid repeated freezing and thawing.
Storage 4 °C/-20 °C
Storage Comment At -20°C for one year. After reconstitution, at 4°C for one month.
It can also be aliquotted and stored frozen at -20 °C for a longer time. Avoid repeated freezing and thawing.
Expiry Date 12 months
Supplier Images
Immunohistochemistry (IHC) image for anti-Mucosal Vascular Addressin Cell Adhesion Molecule 1 (MADCAM1) (AA 173-190), (Middle Region) antibody (ABIN3044076) Anti-MAdCAM1 antibody, IHC(P) IHC(P): Human Appendicitis Tissue
Immunohistochemistry (IHC) image for anti-Mucosal Vascular Addressin Cell Adhesion Molecule 1 (MADCAM1) (AA 173-190), (Middle Region) antibody (ABIN3044076) Anti-MAdCAM1 antibody, IHC(F) IHC(F): Human Placenta Tissue
Western Blotting (WB) image for anti-Mucosal Vascular Addressin Cell Adhesion Molecule 1 (MADCAM1) (AA 173-190), (Middle Region) antibody (ABIN3044076) anti-Mucosal Vascular Addressin Cell Adhesion Molecule 1 (MADCAM1) (AA 173-190), (Middle Region) antibody (Image 3)
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