Article No.: KTSM1341

Specification: 500 μ L

Concentration: 500 μ L agarose microspheres combined with 5.0 mg nanoantibody

Binding ability: 1.0 ml agarose beads can bind about 10 mg of mouse IgG

Product form: 50% agarose microsphere suspension

Host: Alpaca

Target species: mouse

Storage buffer: 20% ethanol solution

Preservative: 0.03% sodium azide

Storage conditions: Refrigerate and store at 4 ° C before opening. Avoid freezing, and the shelf life is 6 months after opening.


"Alpaca heavy chain antibody (HcAb) is a special antibody that exists naturally and consists of only two heavy chains. Alpaca heavy chain antibody (HcAb) binds to an antigen through a variable region (VHH) on the heavy chain, which can independently and stably exist in vitro, and is called a nanoantibody.". The molecular weight of nano antibodies is only 1/10 (about 15KD) of traditional complete antibodies, but they still have complete antigen recognition ability. Generally, complete antibody sequences are obtained through phage screening. Thanks to the tiny structure of nano antibodies, their complete antigen recognition ability, and phage screening technology, nano antibodies exhibit high affinity, high specificity, strong penetration, and ease of modification and expression. Moreover, due to the availability of complete antibody sequences, nano antibodies can be stably produced through in vitro recombinant expression with high quality, effectively avoiding the issue of batch differences between traditional antibodies.

Anti Mouse VHH IP Agarose Beads is a suspension of alpaca nanoantibodies coupled with agarose beads prepared using mouse IgG as an antigen, which is suitable for the isolation and immunoprecipitation experiments of all mouse derived antibodies.

Compared to traditional antibodies or ProteinA/G, this product has several advantages:

*Direct use without coupling proteinA/G and agarose beads

*No heavy and light chain pollution in downstream applications

*Can be cleaned under strict conditions

*5-30min extremely short incubation time

*High yield, stable quality, no batch differences



instructions:

*It is recommended to use a large-diameter pipette to absorb liquid. After opening the vial, it is recommended to seal the bottle cap with a sealing film to prevent evaporation of the buffer solution;

*The beads will sink to the bottom during storage. Before initial use, gently mix the product (do not swirl) to ensure that the agarose beads are evenly suspended in the storage solution.

Immunoprecipitation experiment steps:

1. Preparation of cell lysates:

Plus 50 μ L Anti Mouse Nanobody IP Beads to 500 μ Incubate the lysate in a microcentrifuge tube on ice for 30 minutes. After centrifuging for 3 minutes, transfer the supernatant to a new centrifuge tube.

2. Immunoprecipitation:

Add 5 to a centrifuge tube filled with pre cleaned cell lysate μ G of the first antibody was incubated for 1 hour. Add another 50 μ L Anti Mouse Nanobody IP Beads were incubated on a shaking table for 1 hour. Place the centrifuge tube at 10000 × G Centrifuge for 1 minute. Completely remove the supernatant and use 500 μ Wash 3 times with L cracking buffer (50mM Tris-HCl, pH 8.0; 150mM NaCl; 1% NP-40).

3. SDS-PAGE sample preparation:

After the last wash and aspiration of the supernatant, add 100 μ L Laemmli Buffer (containing 50 mM DTT or 2% 2-mercaptoethanol). Vortex mixing and heating to 90-100 ° C for 10 min, 10000 × G Centrifuge for 3 minutes, collect the supernatant (avoid stirring Anti Rabbit Ig Beads), and conduct SDS-PAGE gel electrophoresis.