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ChromoTek His Fab-Trap® Agarose Kit

The ChromoTek His Fab-Trap® Agarose Kit consists of an anti-His-tag Fab-fragment, which is covalently bound to agarose beads. It also contains lysis, wash, and elution buffers that can be used for the immunoprecipitation of His-tagged proteins from cell extracts of various organisms.

Specificity

6*His, His-Tag (sequence: HHHHHH)

Applications

IP, Co-IP

Type

Fab-fragment

Conjugate

Agarose

Cat no : hfak

Synonyms

His-trap, 6*His Fab-Trap, His-tag Fab-Trap

Validation Data Gallery

  • IP using His Fab-Trap® Agarose (hfa) from untransfected HEK293T (mock) and HEK293T cells transfected with either His-TOM70 or TOM70-His fusion proteins. WB detection of TOM70 protein was also done on samples from the Input (I), Flow-Through (F), and Bound (B) fractions using Anti-TOM70 Polyclonal Antibody (14528-1-AP) and ChromoTek Nano-Secondary alpaca anti-rabbit IgG, CoraLite Plus 488 (srb2GCL488-1).

    IP using His Fab-Trap® Agarose (hfa) from untransfected HEK293T (mock) and HEK293T cells transfected with either His-TOM70 or TOM70-His fusion proteins. WB detection of TOM70 protein was also done on samples from the Input (I), Flow-Through (F), and Bound (B) fractions using Anti-TOM70 Polyclonal Antibody (14528-1-AP) and ChromoTek Nano-Secondary alpaca anti-rabbit IgG, CoraLite Plus 488 (srb2GCL488-1).

  • His Fab-Trap® Agarose (hfa) was used for the Co-IP of AurKinB-His fusion protein and interacting partners from untransfected (mock) and transfected HEK293T cells. Multiplexed WB was done on Input (I), Flow-Through (F), and Bound (B) fractions of the IP. Detection was completed using His-tag Monoclonal Antibody (66005-1-Ig) labeled with FlexAble CoraLite Plus 647 Kit for Mouse IgG (KFA023), CDC37 Monoclonal Antibody (66420-1-Ig) labeled with FLexAble CoraLite Plus 750 Kit for Mouse IgG2a (KFA044), and HSP90AB1 Monoclonal Antibody (67450-1-Ig) labeled with FlexAble CoraLite Plus 488 Kit for Mouse IgG2b (KFA061).

    His Fab-Trap® Agarose (hfa) was used for the Co-IP of AurKinB-His fusion protein and interacting partners from untransfected (mock) and transfected HEK293T cells. Multiplexed WB was done on Input (I), Flow-Through (F), and Bound (B) fractions of the IP. Detection was completed using His-tag Monoclonal Antibody (66005-1-Ig) labeled with FlexAble CoraLite Plus 647 Kit for Mouse IgG (KFA023), CDC37 Monoclonal Antibody (66420-1-Ig) labeled with FLexAble CoraLite Plus 750 Kit for Mouse IgG2a (KFA044), and HSP90AB1 Monoclonal Antibody (67450-1-Ig) labeled with FlexAble CoraLite Plus 488 Kit for Mouse IgG2b (KFA061).

  • IP of His-TOM70 and TOM70-His proteins using either His Fab-Trap Agarose (left) or a competitor resin product (right) from transfected HEK293T cells. The His Fab-Trap pulls down the TOM70 proteins more efficiently and with less background when compared to the competitor.

    IP of His-TOM70 and TOM70-His proteins using either His Fab-Trap Agarose (left) or a competitor resin product (right) from transfected HEK293T cells. The His Fab-Trap pulls down the TOM70 proteins more efficiently and with less background when compared to the competitor.

Product Information

The ChromoTek His Fab-Trap® Agarose Kit consists of an anti-His-tag Fab-fragment, which is covalently bound to agarose beads. It also contains lysis, wash, and elution buffers that can be used for the immunoprecipitation of His-tagged proteins from cell extracts of various organisms.

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

• Fast, reliable & efficient one-step immunoprecipitation

• Ready-to-use

• Low background

• 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 His-tag (also known as hexahistidine or His6-tag) fused to a protein of interest at N-, C- or internal position. Please note that the affinity is highest for a C-terminal fusion (dissociation constant KD of 10 nM for a C-terminal His-tag and 220 nM for an N-terminal His-tag).
Binding Capacity25 ug of recombinant His-tagged protein (~30 kDa) per 25 uL bead slurry
ConjugateAgarose beads; ~90 um (cross-linked 4% agarose beads)
Elution buffer2x SDS-sample buffer (Lämmli), 200 mM glycine pH 2.5, 100 µM His-Peptide
Wash Buffer Compatibility2 M NaCl, 5 mM DTT, 5 mM β-mercaptoethanol, 5 mM TCEP, 2% NP40, 2% Triton X-100, 0.1% SDS, 5 M Urea
TypeFab-fragment
ClassRecombinant
HostMouse
Affinity (KD)10 nM for a C-terminal His-tag and 220 nM for a N-terminal His-tag
Compatibility with mass spectrometryThe His Fab-Trap® is optimized for on-bead digestion. For the application note, please click here:
On-bead digest protocol for mass spectrometry
RRIDAB_3661839
Storage BufferPBS pH 7.4. Preservative: 0.09% sodium azide
Storage ConditionShipped at ambient temperature. Upon receipt store at +4°C. Stable for one year. DO not freeze!

Kit components

Component Description
His Fab-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