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ChromoTek HA-Trap Agarose Kit

The HA-Trap Agarose Kit is a ready-to-use reagent for the IP of HA-tagged proteins. It consists of an anti-HA-tag Nanobody/VHH coupled to agarose beads, along with lysis, wash, and elution buffers to use in the IP process.

Specificity

HA-Tag (sequence YPYDVPDYA)

Applications

IP, Co-IP

Type

Nanobody

Conjugate

Agarose

Cat No : atak

Synonyms

Hemagglutinin tag, HA, HA-tag, HA-tag Trap Agarose

Validation Data Gallery

  • The HA-Trap Agarose Kit was used to immunoprecipitate TOM70-HA fusion protein from either untransfected (mock) HEK293T cells or HEK293T cell transfected with full-length TOM70-HA construct. SDS-PAGE analysis was done on samples from the Input (I), Flow-through (F), Bound (B) fractions.

    The HA-Trap Agarose Kit was used to immunoprecipitate TOM70-HA fusion protein from either untransfected (mock) HEK293T cells or HEK293T cell transfected with full-length TOM70-HA construct. SDS-PAGE analysis was done on samples from the Input (I), Flow-through (F), Bound (B) fractions.

  • Co-IP using HA-Trap Agarose Kit followed by multiplexed WB of TOM70-HA and HSP90 proteins from untransfected (mock) HEK293T cells and HEK293T cells transfected with full-length TOM70-HA construct. WB analysis was done on samples from the Input (I), Flow-through (F) and Bound (B) fractions of the IP. TOM70 Monoclonal Antibody (66593-1-Ig), Multi-rAB CoraLite Plus 488-Goat Anti-Mouse Recombinant Secondary Antibody (RGAM002), HSP90 Polyclonal Antibody (13171-1-AP), and Multi-rAb CoraLite Plus 750-Goat Anti Rabbit Recombinant Secondary Antibody (RGAR006) were used in the WB analysis.

    Co-IP using HA-Trap Agarose Kit followed by multiplexed WB of TOM70-HA and HSP90 proteins from untransfected (mock) HEK293T cells and HEK293T cells transfected with full-length TOM70-HA construct. WB analysis was done on samples from the Input (I), Flow-through (F) and Bound (B) fractions of the IP. TOM70 Monoclonal Antibody (66593-1-Ig), Multi-rAB CoraLite Plus 488-Goat Anti-Mouse Recombinant Secondary Antibody (RGAM002), HSP90 Polyclonal Antibody (13171-1-AP), and Multi-rAb CoraLite Plus 750-Goat Anti Rabbit Recombinant Secondary Antibody (RGAR006) were used in the WB analysis.

Product Information

The HA-Trap Agarose Kit is a ready-to-use reagent for the IP of HA-tagged proteins. It consists of an anti-HA-tag Nanobody/VHH coupled to agarose beads, along with lysis, wash, and elution buffers to use in the IP process.

DescriptionThe HA-Trap Agarose Kit contains HA-Trap agarose, lysis, wash, and elution buffers for efficient immunoprecipitation of HA-tagged proteins.

• Fast, reliable & efficient one-step immunoprecipitation

• Ready-to-use

• No heavy & light antibody chains

• Stable under harsh washing conditions

• Suitable for downstream mass spec analysis

• Works in samples from: mammals, plants, yeast, etc.

ApplicationsIP, Co-IP
Specificity/TargetBinds specifically to the HA-tag (sequence YPYDVPDYA) fused to a protein of interest at N-, C- or internal position. Please note that the affinity is highest for a C-terminal fusion. There is no cross-reactivity to other common peptide tags such as the His6-tag, FLAG-tag, Spot-Tag, V5-tag, Strep-tag, or C-tag (other tags not tested). Background binding to host cell proteins from a range of organisms such as human, mouse and dog cell lines or yeast and plants is low.
Binding Capacity20 ug of recombinant HA-tagged protein (~30 kDa) per 25 uL bead slurry
ConjugateAgarose beads; ~90 um (cross-linked 4% agarose beads)
Elution buffer2x SDS-sample buffer (Lammli)
Wash Buffer Compatibility2M NaCl, 5 mM DTT, 0 mM β-mercaptoethanol, 5 mM TCEP, 2% NP40, 2% Triton X-100, 0% SDS, 2-3 M Urea
TypeNanobody
ClassRecombinant
HostAlpaca
Affinity (KD) 6 nM for C-terminal HA-tags and ca. 180 nM for N-terminal fusions.
Compatibility with mass spectrometryThe HA-Trap is optimized for on-bead digestion. For the application note, please click here:
On-bead digest protocol for mass spectrometry
RRIDAB_3094560
Storage Buffer20% ethanol
Storage ConditionShipped at ambient temperature. Upon receipt store at +4°C. Stable for one year. DO not freeze!

Kit components

Component Description
HA-Trap Agarose20 reactions (500 µl)
Lysis bufferOptimized for cytoplasmic proteins and mammalian cell lysis
RIPA bufferOptimized for nuclear/chromatin proteins and mammalian cell lysis
Wash bufferRemoval of unwanted proteins, peptides, etc.
Dilution bufferDilution of cell lysate
Elution bufferFor acidic elution