Fab fragments of anti fcγ antibody

Manufactured by Jackson ImmunoResearch

Sourced in United States

Fab fragments of anti-Fcγ antibody are laboratory reagents used for immunological applications. They specifically bind to the Fc region of immunoglobulin G (IgG) antibodies. Fab fragments are the antigen-binding portion of the antibody, excluding the Fc region.

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Fab fragments (6 citations)

4 protocols using fab fragments of anti fcγ antibody

Nano-Patterning Proteins on Hydrogels

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This section describes the printing of nano-patterns on “intermediate” cover-glass surface using nano-stamps pre-coated with protein of interest.
Note: For elastic hydrogel micro- and nano-patterning we adopted an “intermediate substrate” technique in which biotin-conjugated protein micro- and nano-patterns were initially printed on “intermediate” cover-glass, then transferred onto hydrogels by cross-linking patterned proteins’ biotin tags to streptavidin-conjugated polyacrylamide (PAA) (Tang et al., 2012 (link)).
Note: To print nano-grids composed of two sets of orthogonally intersecting parallel nanolines, constituted of two different proteins of interest, we use Alexa Fluor™ 568 fluorophore- and biotin-prelabeled anti-collagen rabbit pAb, and Alexa Fluor™ 488 fluorophore- and biotin-prelabeled Fab fragments of anti-Fc_γ antibody (Jackson Immunoresearch).

Tabdanov E.D., Zhovmer A.S., Puram V, & Provenzano P.P. (2020). Engineering Elastic Nano- and Micro-Patterns and Textures for Directed Cell Motility. STAR Protocols, 1(1), 100013.

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Precoating Molds with Fab Fragments

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This section describes the precoating of molding casts with protein of interest for better efficiency of protein-PAA cross-linking.
Note: Weprecoat molding casts with biotinylatedand fluorescenttag-labeled protein of interest(in our case - Fab fragments of anti-Fc_γ antibody (Jackson Immunoresearch) in 0.2 mg/mL PBS solution (See Before You Begin for more details).

Tabdanov E.D., Zhovmer A.S., Puram V, & Provenzano P.P. (2020). Engineering Elastic Nano- and Micro-Patterns and Textures for Directed Cell Motility. STAR Protocols, 1(1), 100013.

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Protein Coating of Nano-Stamps for Printing

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TIMING: 1 or 12 h

This section describes the precoating of nano-stamps with protein of interest for printing on “intermediate” cover-glass.

13.

Place 5 μL droplets of 0.2 mg/mL solution of biotin- and fluorophore-labelled (seesection 2. for more detail) protein of interest (anti-collagen type-1 (AbCam) antibody or Fab fragments of anti-Fc_γ antibody (Jackson Immunoresearch)) atop of the 5✕5 mm or 1✕1 cm square nano-stamps (Figure 2B, step 2).

Note: To ensure an effective stamp surface coating with protein of interest, “sandwich” protein solution droplet between the stamp’s printing surface and pre-baked 15 mm round glass coverslip (See Before You Begin for more details) (Carolina, USA) as shown on Figure 2(step 2).

14.

Incubate the nano-stamps with “sandwiched” droplet of protein of interest solution for 40 minutes at 20-25°C or for ∼8-12 hours at +4°C in wet chamber (See Before You Begin for more details) (Figure 2B, step 3).

PAUSE POINT: Incubation in the wet chamber can be utilized as a pause point, 40 min or 8-12 hours.

15.

After the coating session the nano-stamps were gently rinsed in deionized milli-Q water (Q-water) and gently dried under the jet of filtered air, nitrogen or argon, and then used for nano-printing.

Tabdanov E.D., Zhovmer A.S., Puram V, & Provenzano P.P. (2020). Engineering Elastic Nano- and Micro-Patterns and Textures for Directed Cell Motility. STAR Protocols, 1(1), 100013.

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Protein-Assisted Microfluidic Device Fabrication

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TIMING: 1 or 12 h

This section describes the precoating of molding casts with protein of interest for better efficiency of protein-PAA cross-linking.

Note: We precoat molding casts with biotinylated and fluorescent tag-labeled protein of interest (in our case - Fab fragments of anti-Fc_γ antibody (Jackson Immunoresearch) in 0.2 mg/mL PBS solution (See Before You Begin for more details).

Put a 5-7 μL droplet of protein of interest solution atop of the hPDMS mold and cover it by the round 15 mm pre-baked cover-glass (Carolina Biological Supply Company) (See Before You Begin for more details) in the “sandwich” fashion (Figure 1C, steps 6-7) to economize the use of protein solution and to ensure its even distribution. Incubate at 20-25°C for 40 minutes or at +4°C for 8-12 hours in a wet chamber (20✕160 mm Petri dish with Q-water-wetted tissue or cotton balls).

After incubation, gently rinse the casting mold in deionized Q-water and gently dry it under the jet of filtered air, nitrogen or argon (Figure 1C), use immediately.

Tabdanov E.D., Zhovmer A.S., Puram V, & Provenzano P.P. (2020). Engineering Elastic Nano- and Micro-Patterns and Textures for Directed Cell Motility. STAR Protocols, 1(1), 100013.

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