β-Actin Loading Control for Westerns using Nitrocellulose Membranes; 3 Lanes loaded with mouse muscle homogenate (5 ug total protein, 10 ug total protein, 20 ug total protein); 1' antibody -- ACT (1:1000); 2' antibody -- HRP Goat anti-chicken IgY (Aves Labs, 1:2000)
from Aves Labs
Anti-Actin (Nitrocellulose Membrane Loading Control) Antibody (ACT)
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Our anti-actin antibody (made against rabbit muscle actin) recognizes mouse, rat and human actin proteins in SDS-polyacrylamide gel transfers onto nitrocellulose membranes. This loading control assures that similar amounts of total proteins were added to each of the lanes in the gels. Note that this product does not work in transfers using PVDF membranes.
Volume: 100 µL (ACT-1010) or 400 µL (ACT-1040)
Concentration: 10 mg/mL
Clonality: Polyclonal
Host Species: Chicken
Species Reactivity: Human, Mouse, Rat
Applications: WB
Antibody Registry ID (RRID): AB_2313504
Physical State: Liquid
Buffer: Sodium phosphate (10 mM, pH 7.2) buffered isotonic saline (0.9%, w/v), glycerol (50%, v/v), with sodium azide (0.02%, w/v) as an anti-microbial agent.
Production Notes: Chickens were immunized with Actin purified from rabbit muscle. After repeated injections, immune eggs were collected, and the IgY fractions were purified from the yolks. These IgY fractions were then affinity-purified using a peptide column. The final product contains 50% glycerol (v/v), purified anti-Actin IgY fraction (10 mg/ml) spiked with 25 µg/ml of affinity purified anti-Actin IgY, and then filter-sterilized.
Validation and Application Notes
Molecular Weight: 42 kDa
Quality Control
This anti-Actin IgY preparation was analyzed by Western Blot analysis (at a dilution of 1:2000) using HRP-labeled goat anti-chicken IgY (1:5000 dilution, Aves Labs Cat. # H-1004) as the secondary reagent.
Storage
Store at -20˚C in the dark. Under these conditions, the antibodies should have a shelf life of at least 12 months (provided they remain sterile). Do not freeze antibodies at -80˚C, unless you want to store them for longer periods of time. Note that each time an antibody preparation is frozen about half its binding activity is lost.
NOTE
Aves Labs products are intended for use as research laboratory reagents. They are not intended for use as diagnostic or therapeutic reagents in humans.
Citations
- Tian S, Muneeruddin K, Choi MY, Tao L, Bhuiyan RH, Ohmi Y, Furukawa K, Furukawa K, Boland S, Shaffer SA, Adam RM, Dong M. (2018), 'Genome-wide CRISPR screens for Shiga toxins and ricin reveal Golgi proteins critical for glycosylation.' Plos Biology. 10.1371/journal.pbio.2006951.