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ChromoTek HA-Trap Magnetic Particles M-270

The ChromoTek HA-Trap Magnetic Particles M-270 consists of an anti-HA-tag Nanobody/VHH, which is coupled to magnetic particles. It can be used for the immunoprecipitation of HA-tagged proteins from cell extracts of various organisms such as humans, mice, dogs, plants, and yeast. It is highly recommended when very large proteins/complexes are being investigated.

Specificity

HA-Tag (sequence YPYDVPDYA)

Applications

IP, Co-IP

Type

Nanobody

Conjugate

Magnetic Particles M-270

Cat No : atd

Synonyms

Hemagglutinin tag, HA, HA-tag, HA-tag Trap Magnetic Particles

Validation Data Gallery

  • The HA-Trap Magnetic Particles M-270 (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 Magnetc Particles M-270 results in cleaner, 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 Magnetic Particles M-270 (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 Magnetc Particles M-270 results in cleaner, 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.

  • WB detection of TOM70-HA fusion protein following immunoprecipitation with HA-Trap Magnetic Particles M-270 from either untransfected (mock) HEK293T cells or HEK293T cells transfected with full-length TOM70-HA construct. Samples from the Input (I), Flow-Through (F), and Bound (B) fractions were used in the WB analysis. Detection was completed using TOM70 Monoclonal Antibody (66593-1-Ig) and Multi-rAb HRP-Goat Anti-Mouse Recombinant Secondary Antibody (RGAM001).

    WB detection of TOM70-HA fusion protein following immunoprecipitation with HA-Trap Magnetic Particles M-270 from either untransfected (mock) HEK293T cells or HEK293T cells transfected with full-length TOM70-HA construct. Samples from the Input (I), Flow-Through (F), and Bound (B) fractions were used in the WB analysis. Detection was completed using TOM70 Monoclonal Antibody (66593-1-Ig) and Multi-rAb HRP-Goat Anti-Mouse Recombinant Secondary Antibody (RGAM001).

  • The HA-Trap Magnetic Particles M-270 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 Magnetic Particles M-270 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 Magnetic Particles M-270 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 utizling 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 timers resulting in co-elution of endogenous PCNA proteins with HA-tagged PCNA.

    The HA-Trap Magnetic Particles M-270 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 utizling 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 timers resulting in co-elution of endogenous PCNA proteins with HA-tagged PCNA.

Product Information

The ChromoTek HA-Trap Magnetic Particles M-270 consists of an anti-HA-tag Nanobody/VHH, which is coupled to magnetic particles. It can be used for the immunoprecipitation of HA-tagged proteins from cell extracts of various organisms such as humans, mice, dogs, plants, and yeast. It is highly recommended when very large proteins/complexes are being investigated.

DescriptionImmunoprecipitation of HA-fusion proteins with anti-HA Nanobody conjugated to magnetic particles M-270. Highly recommended when very large proteins/complexes are being investigated.

• Fast, reliable & efficient one-step immunoprecipitation without size limitation

• Ready-to-use

• No heavy & light antibody chains

• Stable under harsh washing conditions

• Suitable for automated and high throughput applications

• 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 Capacity5 ug of recombinant HA-tagged protein (~30 kDa) per 25 uL bead slurry
ConjugateMagnetic Particles M-270, size: 2.8 µm
Elution buffer2x SDS-sample buffer (Lammli)
Wash Buffer Compatibility<1 M NaCl, 5 mM DTT, 5 mM β-mercaptoethanol, 5 mM TCEP, 2% NP40, 2% Triton X-100, 0 % SDS, 1 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_3094562
Storage BufferPBS with 0.09% sodium azide
Storage ConditionShipped at ambient temperature. Upon receipt store at +4°C. Stable for one year. DO not freeze!