E.coli-derived human ILKAP recombinant protein (Position: E90-H392). Human ILKAP shares 98% and 98.7% amino acid (aa) sequence identity with mouse and rat ILKAP, respectively.
Conjugation:
Unconjugated
Alternative Names:
Ly6/PLAUR domain-containing protein 3, GPI-anchored metastasis-associated protein C4.4A homolog, Matrigel-induced gene C4 protein, MIG-C4, LYPD3, C4.4A
Anti-LYPD3 Antibody. Tested in ELISA, IF, IHC, WB, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat.
Clonality:
Polyclonal
Concentration:
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Western blot, 0.25-0.5 µg/ml, Human, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 1-2 µg/ml, Human, Rat Immunofluorescence, 5 µg/ml, Human Flow Cytometry (Fixed), 1-3 µg/1x10 6 cells, Human ELISA, 0.1-0.5 µg/ml, -
Flow Cytometry analysis of RT4 cells using anti-LYPD3 antibody. Overlay histogram showing RT4 cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-LYPD3 Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
IF analysis of LYPD3 using anti-LYPD3 antibody. LYPD3 was detected in a paraffin-embedded section of human skin cancer 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 5 µg/mL rabbit anti-LYPD3 Antibody overnight at 4C. Cy3 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
IHC analysis of LYPD3 using anti-LYPD3 antibody. LYPD3 was detected in a paraffin-embedded section of human skin cancer 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 2 µg/ml rabbit anti-LYPD3 Antibody overnight at 4C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
IHC analysis of LYPD3 using anti-LYPD3 antibody. LYPD3 was detected in a paraffin-embedded section of rat stomach 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 2 µg/ml rabbit anti-LYPD3 Antibody overnight at 4C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
Western blot analysis of LYPD3 using anti-LYPD3 antibody. 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 30 ug of sample under reducing conditions. Lane 1: human RT4 whole cell lysates, Lane 2: human A431 whole cell lysates, Lane 3: rat stomach tissue lysates, Lane 4: mouse stomach tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-LYPD3 antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4C, 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 with Tanon 5200 system. A specific band was detected for LYPD3 at approximately 75 kDa. The expected band size for LYPD3 is at 36 kDa.
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