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Product Info Summary
| SKU: | A02518-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-Milk Fat Globule 1/Mfge8 Antibody Picoband®
SKU/Catalog Number
A02518-1
Size
100 μg/vial
Form
Lyophilized
Description
MFGE8 (MFG-E8) is a phosphatidylserine-binding opsonin that bridges apoptotic cells to phagocyte integrins (e.g., αvβ3/αvβ5), supporting efferocytosis and inflammation resolution; deficiency models show lupus-like autoimmunity, consistent with a clearance-to-tolerance link (context dependent). Assay context: mouse/rat-reactive antibody tested for ELISA, flow cytometry, IHC, and WB. Often interpreted with macrophage-lineage markers such as CD163 and resolution mediators like ANXA1 when studying apoptotic clearance vs persistent inflammation (putative co-panel).
Storage & Handling
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.
Cite This Product
Anti-Milk Fat Globule 1/Mfge8 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02518-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived mouse Milk Fat Globule 1/Mfge8 recombinant protein (Position: A23-C463). Mouse Mfge8 shares 86.2% amino acid (aa) sequence identity with rat Mfge8.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A02518-1 is reactive to Mfge8 in Mouse, Rat
Observed Molecular Weight
65-70 kDa
Calculated molecular weight
51.2 kDa
Background of Mfge8
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.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A02518-1 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Mouse
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: rat kidney tissue, rat liver tissue, mouse NIH/3T3 whole cell
IHC: mouse kidney tissue
FCM: HEPA1-6 cell
Validation Images & Assay Conditions
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Western blot analysis of Milk Fat Globule 1/Mfge8 using anti-Milk Fat Globule 1/Mfge8 antibody (A02518-1).
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 lysates,
Lane 2: rat liver tissue lysates,
Lane 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 # A02518-1) at 0.5 μg/mL overnight at 4°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 # EK1002) 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.
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IHC analysis of Milk Fat Globule 1/Mfge8 using anti-Milk Fat Globule 1/Mfge8 antibody (A02518-1).
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 (A02518-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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Flow Cytometry analysis of HEPA1-6 cells using anti-Milk Fat Globule 1/Mfge8 antibody (A02518-1).
Overlay histogram showing HEPA1-6 cells stained with A02518-1 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Milk Fat Globule 1/Mfge8 Antibody (A02518-1, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-Milk Fat Globule 1/Mfge8 Antibody Picoband® (A02518-1)
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