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ChromoTek HA-Trap Agarose

The ChromoTek HA-Trap Agarose consists of an anti-HA tag Nanobody/VHH, which is coupled to agarose beads. It can be used for the immunoprecipitation of HA-tagged proteins from cell extracts of various organisms such as humans, mice, dogs, yeast, and plants.

Specificity

HA-Tag (sequence YPYDVPDYA)

Applications

IP, Co-IP

Type

Nanobody

Conjugate

Agarose

Cat no : ata

Synonyms

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

Validation Data Gallery

  • The HA-Trap Agarose (left) and a competitor resin (right) were used to immunoprecipitate TOM70-HA fusion protein from either untransfected (mock) HEK293T cells or HEK293T cell transfected with full-length TOM70-HA construct. Immunoprecipitation with HA-Trap Agarose results in clean, single-band pulldowns without any heavy and light chain contamination. SDS-PAGE analysis was done on samples from the Input (I), Flow-through (F), Bound (B) fractions.

    The HA-Trap Agarose (left) and a competitor resin (right) were used to immunoprecipitate TOM70-HA fusion protein from either untransfected (mock) HEK293T cells or HEK293T cell transfected with full-length TOM70-HA construct. Immunoprecipitation with HA-Trap Agarose results in clean, single-band pulldowns without any heavy and light chain contamination. SDS-PAGE analysis was done on samples from the Input (I), Flow-through (F), Bound (B) fractions.

  • Co-IP using HA-Trap Agarose 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 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.

  • The HA-Trap Agarose was used to immunoprecipitate HA-PCNA and PCNA-HA proteins from transfected HEK293T cells. WB analysis was done on samples from the Input (I), Flow-Through (F), and Bound (B) fractions of the IP using PCNA Monclonal Antibody (60097-1-Ig) and Multi-rAb HRP-Goat Anti-Mouse Recombinant Secondary Antibody (RGAM001). The HA-Trap is succesful in pulling down HA-tagged PCNA regardless of whether the tag is fused to the N- or C-terminal. Note: PCNA forms trimers, resulting in co-elution of endogenous PCNA with HA-tagged PCNA.

    The HA-Trap Agarose was used to immunoprecipitate HA-PCNA and PCNA-HA proteins from transfected HEK293T cells. WB analysis was done on samples from the Input (I), Flow-Through (F), and Bound (B) fractions of the IP using PCNA Monclonal Antibody (60097-1-Ig) and Multi-rAb HRP-Goat Anti-Mouse Recombinant Secondary Antibody (RGAM001). The HA-Trap is succesful in pulling down HA-tagged PCNA regardless of whether the tag is fused to the N- or C-terminal. Note: PCNA forms trimers, resulting in co-elution of endogenous PCNA with HA-tagged PCNA.

  • The HA-Trap Agarose was used to immunoprecipitate HA-PCNA fusion protein from HEK293T cells. HA-PCNA protein was released from the trap through a two-step competitive elution with HA-peptide (ap). Samples from the Input (I), Flow-Through (F), 1st elution (E1), 2nd elution (E2), and residual (R) fractions were analyzed through WB. PCNA Monoclonal Antibody (60097-1-Ig) and Multi-rAb HRP-Goat Anti-Mouse Recombinant Secondary Antibody (RGAM001) were used in the WB analysis. Note: PCNA forms trimers resulting in co-elution of endogenous PCNA proteins with HA-tagged PCNA.

    The HA-Trap Agarose was used to immunoprecipitate HA-PCNA fusion protein from HEK293T cells. HA-PCNA protein was released from the trap through a two-step competitive elution with HA-peptide (ap). Samples from the Input (I), Flow-Through (F), 1st elution (E1), 2nd elution (E2), and residual (R) fractions were analyzed through WB. PCNA Monoclonal Antibody (60097-1-Ig) and Multi-rAb HRP-Goat Anti-Mouse Recombinant Secondary Antibody (RGAM001) were used in the WB analysis. Note: PCNA forms trimers resulting in co-elution of endogenous PCNA proteins with HA-tagged PCNA.

Product Information

The ChromoTek HA-Trap Agarose consists of an anti-HA tag Nanobody/VHH, which is coupled to agarose beads. It can be used for the immunoprecipitation of HA-tagged proteins from cell extracts of various organisms such as humans, mice, dogs, yeast, and plants.

DescriptionImmunoprecipitation of HA-tagged proteins with anti-HA-tag Nanobody conjugated to agarose beads.

• 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 (~30kDa) 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!
Size25ul/reactions (eg：20rxns=500ul slurry)

Publications

ApplicationTitle
CoIP

Am J Hum Genet

Bi-allelic KICS2 mutations impair KICSTOR complex-mediated mTORC1 regulation, causing intellectual disability and epilepsy

Authors - Rebecca Buchert
View Article