Anti-Milk Fat Globule 1/Mfge8 Antibody Picoband™

Product Name

Anti-Milk Fat Globule 1/Mfge8 Antibody Picoband™

View all MFG-E8 Antibodies

SKU/Catalog Number

A02518-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Milk Fat Globule 1/Mfge8 Antibody Picoband™ catalog # A02518-1. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Mouse, Rat.

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.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A02518-1 is reactive to MFGE8 in Mouse, Rat

Applications

A02518-1 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

65-70 kDa

Calculated molecular weight

43.105kDa

Background of MFG-E8

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.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

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, 1-3μg/1x10⁶ cells, Mouse
Direct ELISA, 0.1-0.5μg/ml, Mouse

Validation Images & Assay Conditions

Click image to see more details

Figure 1. 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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Figure 2. 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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Figure 3. 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 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 (Red line) was also used as a control.

Gene/Protein Information For MFGE8 (Source: Uniprot.org, NCBI)

Alternative Names

BA46; Breast epithelial antigen BA46; EDIL1; hP47; HsT19888; lactadherin; Lactahedrin; Medin; MFG1; MFGE8; MFG-E8; MFGM; milk fat globule-EGF factor 8 protein; Milk fat globule-EGF factor 8; O-acetyl disialoganglioside synthase; OAcGD3S; SED1; SPAG10; sperm associated antigen 10; sperm surface protein hP47 MFGE8 BA46, EDIL1, HMFG, HsT19888, MFG-E8, MFGM, OAcGD3S, SED1, SPAG10, hP47 milk fat globule EGF and factor V/VIII domain containing lactadherin|O-acetyl disialoganglioside synthase|breast epithelial antigen BA46|medin|milk fat globule-EGF factor 8 protein|sperm associated antigen 10|sperm surface protein hP47

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