LY6C Antibody

• Catalog Number: orb654438
• Category: Antibodies
• Description: LY6C Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, IHC, WB
• Reactivity: Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived mouse LY6C recombinant protein (Position: L27-G109).
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.1-0.25μg/ml, Mouse Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Mouse, Rat Flow Cytometry, 1-3μg/1x106 cells, Mouse Direct ELISA, 0.1-0.5μg/ml, Mouse
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 14 kDa
• UniProt ID: P0CW02
• Storage: 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.
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
• Expiration Date: 12 months from date of receipt.

References

Wanyuji Wang 1, Xueling Zheng 1, Hehe Wang 1, Bin Zuo 2, Sisi Chen 2, Jiao Li Mechanical Unloading Promotes Osteoclastic Differentiation and Bone Resorption by Modulating the MSC Secretome to Favor Inflammation Cell Transplant, 33, 9636897241236584 (2024)

PubMed: 38501500

Applications

FC

Reactivity

Rat

Western blot analysis of LY6C using anti-LY6C antibody (orb654438). 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 50ug of sample under reducing conditions. Lane 1: mouse RAW264.7 whole cell lysates, Lane 2: mouse ANA-1 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-LY6C antigen affinity purified polyclonal antibody (Catalog # orb654438) at 0.25 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 # orb90503) with Tanon 5200 system. A specific band was detected for LY6C at approximately 14KD. The expected band size for LY6C is at 14KD.

IHC analysis of LY6C using anti-LY6C antibody (orb654438). LY6C was detected in paraffin-embedded section of rat spleen 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 1μg/ml rabbit anti-LY6C Antibody (orb654438) 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 # orb90444) with DAB as the chromogen.

Flow Cytometry analysis of mouse spleen tissue using anti-LY6C antibody (orb654438). Overlay histogram showing mouse spleen tissue stained with orb654438 (Blue line). The tissue was blocked with 10% normal goat serum. And then incubated with rabbit anti-LY6C Antibody (orb654438, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

IHC analysis of LY6C using anti-LY6C antibody (orb654438). LY6C was detected in paraffin-embedded section of mouse spleen 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 1μg/ml rabbit anti-LY6C Antibody (orb654438) 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 # orb90444) with DAB as the chromogen.