ChromoTek BSA Depletion Agarose Kit

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Validation Data Gallery

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  BSA depletion from various antibodies. Proteintech BSA Depletion Agarose was validated with antibodies from various suppliers and with varying BSA content (from 0 – 0.1%). IgGs run at ~150 kDa (non-reducing SDS PAGE), BSA runs at ~50 kDa. Comparing the Input (In) vs. output (Out) lanes consistently shows successful removal of BSA with minimal residual BSA in the output fraction. Poly = Polyclonal, Ms = Mouse, Rb = Rabbit, Ch, Sp = Sheep, Ch = Chicken, Rt = Rat

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  BSA depletion benchmarking.
  Proteintech BSA Depletion Beads (Lane 4) were evaluated for BSA depletion efficiency and compared with competitor products. The BSA reference (Lane 2) contains 0.1% BSA and is contaminated with bovine IgG (which is normal). The input antibody contains 1% BSA (lane 3). Proteintech BSA Depletion Beads removes approximately 90% of the BSA. In contrast, competitor products leave high residual BSA levels, thus also failing to remove bovine IgG that interferes with accurate quantification of the target IgG (Lanes 5 and 6). Other competitor products also exhibit reduced recovery of the target IgG (Lane 7), most evident from the Western blot result. Western blot analysis using an anti-rabbit IgG antibody confirms efficient recovery of the target IgG following Proteintech BSA Depletion Beads treatment, with no evident loss of the target IgG (here: rabbit IgG).

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  Immunoprecipitation of serum albumin from different species using BSA Depletion Agarose. BSA Depletion beads were used to test for cross reactivity to albumnin from other species. As expected, strong cross-reactivity to albumin from closely related species (bovid family: sheep, goat) can be observed. Other serum albumin species do not bind to the beads (e.g., HSA from humans)

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  BSA depletion benchmarking.
  Proteintech BSA Depletion Agarose (Lane 3) was evaluated for BSA depletion efficiency and compared with competitor products. The starting antibody (Lane 2) contains 0.01% BSA. Proteintech BSA Depletion Beads remove approximately 95% of the BSA. In contrast, competitor products partly leave higher residual BSA levels. Other competitor products also exhibit reduced recovery of the target IgG (Lane 6).

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  Reference table for bead usage per depletion.
  Table 1 shows the recommended bead volume for each antibody solution volume (top row) and corresponding BSA concentration (%) (first column).
  Table 2 shows the expected number of depletions for each unit size (1 mL, 2 mL, and 5 mL), based on the antibody solution volume and BSA concentration, using the recommended bead volumes specified in Table 1.

ChromoTek BSA Depletion Agarose consists of an anti-BSA VHH (Nanobody), which is coupled to agarose beads. It can be used for fast & efficient removal of BSA from antibodies that are supplied with BSA containing buffers.
Note: BSA Depletion Agarose Kits contain spin columns (cat.# sct).

Cat No. MG-3232AK

Reactivity

BSA

Applications

Depletion (Removal of BSA from Antibody)

Conjugate

Agarose

Synonyms

BSA Depletion, BSA removal

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Product Information

ChromoTek BSA Depletion Agarose consists of an anti-BSA VHH (Nanobody), which is coupled to agarose beads. It can be used for fast & efficient removal of BSA from antibodies that are supplied with BSA containing buffers.
Note: BSA Depletion Agarose Kits contain spin columns (cat.# sct).

• Description: BSA depletion agarose facilitates fast & easy removal of BSA from antibodies with high antibody recovery. BSA is often added to antibodies for stabilization but may interfere with conjugation methods and other downstream applications. ; ; General Notes ; • BSA Depletion Agarose is a high-affinity binding resin binding BSA to deplete it from an antibody solution. The antibody of interest will flow through. ; • BSA Depletion Agarose is compatible with antibodies from a wide range of species such as rabbit, mouse, rat, chicken, sheep etc. ; • BSA Depletion Agarose is also compatible with typical antibody buffer compounds, including glycerol up to 50%. ; • BSA concentrations in antibodies can vary from 0.01% to 1%. Our recommendation for usage: ; ;

  ▪ Use 50 µl BSA Depletion Agarose (slurry) for 100 µl of antibody supplemented with ≤ 0.1% BSA.;

  ▪ Use 200 µl beads (slurry) for 100 µl antibody solutions with 0.5% BSA.;

  ▪ Use 300 µl beads (slurry) for 100 µl antibody solutions with 1% BSA.

  ; Note: We do not recommend using less than 50 µl to avoid significant loss of antibody due to bead handling issues. ; Bead volumes can be scaled up proportionally to the antibody volume. ; ; • Important note: Commercial BSA preparations often contain bovine serum proteins, including bovine immunoglobulins (IgG). As a result, the recovered IgG may appear lower in quantity, when in fact the purification has removed contaminating bovine IgG and yielded a cleaner preparation of the target IgG.; ; A detailed protocol is provided in the documentation section below.
• Applications: Depletion (Removal of BSA from Antibody)
• Bead concentration: 50% Slurry
• Resin Type: Agarose beads; bead size: ~ 90 µm (cross-linked 4 % agarose beads)
• Type: VHH
• Class: Recombinant - Animal free production
• Immunogen: BSA
• Affinity: 3 nM
• RRID: AB_3753744
• Storage Buffer: 20% Ethanol
• Storage Condition: Upon arrival store at +4°C / do not freeze!
• Shipping: Ambient Temp

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