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Anti-AFM Antibody

Catalog Number: orb669154

DispatchUsually dispatched within 3-4 weeks
$ 210.00
Catalog Numberorb669154
CategoryAntibodies
DescriptionAnti-AFM Antibody. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Human.
Species/HostRabbit
ClonalityPolyclonal
Tested applicationsELISA, FC, IHC, WB
ReactivityHuman
IsotypeRabbit IgG
ImmunogenE.coli-derived human AFM recombinant protein (Position: D41-E584).
Antibody TypePrimary Antibody
ConcentrationAdding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
Form/AppearanceLyophilized
ConjugationUnconjugated
MW87 kDa
UniProt IDP43652
StorageMaintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Alternative namesAgouti-related protein; Agrp; Agrt; Art
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NoteFor research use only
Application notesWestern blot, 0.1-0.25μg/ml, Human Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human ELISA, 0.1-0.5μg/ml, -. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
Expiration Date12 months from date of receipt.
Anti-AFM Antibody

Flow Cytometry analysis of HL-60 cells using anti-AFM antibody. Overlay histogram showing HL-60 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-AFM Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Anti-AFM Antibody

IHC analysis of AFM using anti-AFM antibody. AFM was detected in paraffin-embedded section of human liver 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-AFM Antibody 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) with DAB as the chromogen.

Anti-AFM Antibody

IHC analysis of AFM using anti-AFM antibody. AFM was detected in paraffin-embedded section of human ovarian 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-AFM Antibody 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) with DAB as the chromogen.

Anti-AFM Antibody

Western blot analysis of AFM using anti-AFM 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 50 ug of sample under reducing conditions. Lane 1: human plasma 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-AFM antigen affinity purified polyclonal antibody 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 with Tanon 5200 system. A specific band was detected for AFM at approximately 87 KD. The expected band size for AFM is at 87 KD.

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