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Western Blot (WB) (Figure 1. Western blot analysis of Milk Fat Globule 1/Mfge8 using anti-Milk Fat Globule 1/Mfge8 antibody (MBS1753533).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: rat kidney tissue lysatesLane 2: rat liver tissue lysatesLane 3: mouse NIH/3T3 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-Milk Fat Globule 1/Mfge8 antigen affinity purified polyclonal antibody (Catalog # MBS1753533) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for Milk Fat Globule 1/Mfge8 at approximately 65-70KD. The expected band size for Milk Fat Globule 1/Mfge8 is at 65-70KD. )

Rabbit anti-Mouse, Rat Milk Fat Globule 1/Mfge8 Polyclonal Antibody | anti-Mfge8 antibody

Anti-Milk Fat Globule 1/Mfge8 Antibody

Gene Names
Mfge8; P47; MP47; Mfgm; SED1; MFG-E8; AA408458; AI325141; lactadherin
Reactivity
Mouse, Rat
Applications
Western Blot, Immunohistochemistry, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
Milk Fat Globule 1/Mfge8; Polyclonal Antibody; Anti-Milk Fat Globule 1/Mfge8 Antibody; Mfge8; actadherin; MFGM; Milk fat globule-EGF factor 8; MFG-E8; SED1; Sperm surface protein SP47; MP47; milk fat globule EGF and factor V/VIII domain containing; anti-Mfge8 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Rabbit IgG polyclonal antibody for Milk Fat Globule 1/Mfge8 detection.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Applicable Applications for anti-Mfge8 antibody
Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA)
Application Notes
WB: 0.25-0.5ug/ml|Mouse, Rat|
IHC-P: 2-5ug/ml|Mouse|
FC/FACS/FCM: 1-3ug/1x106 cells|Mouse|
Direct ELISA: 0.1-0.5ug/ml|Mouse|
Immunogen
E Coli-derived mouse Milk Fat Globule 1/Mfge8 recombinant protein (Position: A23-C463).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Recommended Detection Systems
Recommended Detection Systems
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. Western blot analysis of Milk Fat Globule 1/Mfge8 using anti-Milk Fat Globule 1/Mfge8 antibody (MBS1753533).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: rat kidney tissue lysatesLane 2: rat liver tissue lysatesLane 3: mouse NIH/3T3 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-Milk Fat Globule 1/Mfge8 antigen affinity purified polyclonal antibody (Catalog # MBS1753533) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for Milk Fat Globule 1/Mfge8 at approximately 65-70KD. The expected band size for Milk Fat Globule 1/Mfge8 is at 65-70KD. )

Western Blot (WB) (Figure 1. Western blot analysis of Milk Fat Globule 1/Mfge8 using anti-Milk Fat Globule 1/Mfge8 antibody (MBS1753533).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: rat kidney tissue lysatesLane 2: rat liver tissue lysatesLane 3: mouse NIH/3T3 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-Milk Fat Globule 1/Mfge8 antigen affinity purified polyclonal antibody (Catalog # MBS1753533) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for Milk Fat Globule 1/Mfge8 at approximately 65-70KD. The expected band size for Milk Fat Globule 1/Mfge8 is at 65-70KD. )

Immunohistochemistry (IHC)

(Figure 2. IHC analysis of Milk Fat Globule 1/Mfge8 using anti-Milk Fat Globule 1/Mfge8 antibody (MBS1753533).Milk Fat Globule 1/Mfge8 was detected in paraffin-embedded section of mouse kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Milk Fat Globule 1/Mfge8 Antibody (MBS1753533) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )

Immunohistochemistry (IHC) (Figure 2. IHC analysis of Milk Fat Globule 1/Mfge8 using anti-Milk Fat Globule 1/Mfge8 antibody (MBS1753533).Milk Fat Globule 1/Mfge8 was detected in paraffin-embedded section of mouse kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Milk Fat Globule 1/Mfge8 Antibody (MBS1753533) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )

Flow Cytometry (FC/FACS)

(Figure 3. Flow Cytometry analysis of HEPA1-6 cells using anti-Milk Fat Globule 1/Mfge8 antibody (MBS1753533).Overlay histogram showing HEPA1-6 cells stained with MBS1753533 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Milk Fat Globule 1/Mfge8 Antibody (MBS1753533, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS) (Figure 3. Flow Cytometry analysis of HEPA1-6 cells using anti-Milk Fat Globule 1/Mfge8 antibody (MBS1753533).Overlay histogram showing HEPA1-6 cells stained with MBS1753533 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Milk Fat Globule 1/Mfge8 Antibody (MBS1753533, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Related Product Information for anti-Mfge8 antibody
MFGE8(Milk Fat Globule-Egf Factor 8), also called as Lactadherin or SED1, is a protein which in humans is encoded by the MFGE8 gene. Mfge8 is secreted protein found in vertebrates, including mammals as well as birds. By fluorescence in situ hybridization, Collins et al. (1997) mapped the MFGE8 gene to chromosome 15q25. Hanayama et al. (2002) found that MFGE8 is a factor that links apoptotic cells to phagocytes. MFGE8 specifically bound to apoptotic cells by recognizing aminophospholipids such as phosphatidylserine. MFGE8, when engaged by phospholipids, bound to cells via its RGD motif. It bound particularly strongly to cells expressing alpha-V-beta-3 integrin. Bu et al. (2007) showed that Mfge8 was expressed in intestinal lamina propria macrophages in mice. Using a wound-healing assay, they showed that Mfge8 promoted migration of intestinal epithelial cells through a PKC-epsilon(PRKCE)-dependent mechanism.
References
1. Bu, H. -F., Zuo, X. -L., Wang, X., Ensslin, M. A., Koti, V., Hsueh, W., Raymond, A. S., Shur, B. D., Tan, X. -D. Milk fat globule-EGF factor 8/lactadherin plays a crucial role in maintenance and repair of murine intestinal epithelium. J. Clin. Invest. 117: 3673-3683, 2007.
2. Collins, C., Nehlin, J. O., Stubbs, J. D., Kowbel, D., Kuo, W. -L., Parry, G. Mapping of a newly discovered human gene homologous to the apoptosis associated-murine mammary protein, MFG-E8, to chromosome 15q25. Genomics 39: 117-118, 1997.
3. Hanayama, R., Tanaka, M., Miwa, K., Shinohara, A., Iwamatsu, A., Nagata, S. Identification of a factor that links apoptotic cells to phagocytes. Nature 417: 182-187, 2002.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
UniProt Accession #
Molecular Weight
47,169 Da
NCBI Official Full Name
Lactadherin
NCBI Official Synonym Full Names
milk fat globule-EGF factor 8 protein
NCBI Official Symbol
Mfge8
NCBI Official Synonym Symbols
P47; MP47; Mfgm; SED1; MFG-E8; AA408458; AI325141; lactadherin
NCBI Protein Information
lactadherin; EGF/factor VIII; sperm surface protein SP47; milk fat globule glycoprotein
UniProt Protein Name
Lactadherin
Protein Family
UniProt Gene Name
Mfge8
UniProt Synonym Gene Names
MFG-E8; MP47
UniProt Entry Name
MFGM_MOUSE

Uniprot Description

MFGE8: Plays an important role in the maintenance of intestinal epithelial homeostasis and the promotion of mucosal healing. Promotes VEGF-dependent neovascularization. Contributes to phagocytic removal of apoptotic cells in many tissues. Specific ligand for the alpha-v/beta-3 and alpha-v/beta-5 receptors. Also binds to phosphatidylserine-enriched cell surfaces in a receptor-independent manner. Zona pellucida-binding protein which may play a role in gamete interaction. Binds specifically to rotavirus and inhibits its replication. 3 isoforms of the human protein are produced by alternative splicing.

Protein type: Secreted; Secreted, signal peptide

Cellular Component: extrinsic to plasma membrane; extracellular matrix; extracellular space; membrane; extracellular region; vesicle; external side of plasma membrane

Molecular Function: integrin binding; phosphatidylethanolamine binding; phosphatidylserine binding

Biological Process: single fertilization; positive regulation of phagocytosis; phagocytosis, recognition; angiogenesis; phagocytosis, engulfment; cell adhesion

Research Articles on Mfge8

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Product Notes

The Mfge8 mfge8 (Catalog #AAA1753533) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-Milk Fat Globule 1/Mfge8 Antibody reacts with Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's Milk Fat Globule 1/Mfge8 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Mouse, Rat| IHC-P: 2-5ug/ml|Mouse| FC/FACS/FCM: 1-3ug/1x106 cells|Mouse| Direct ELISA: 0.1-0.5ug/ml|Mouse|. Researchers should empirically determine the suitability of the Mfge8 mfge8 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Milk Fat Globule 1/Mfge8, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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