ChromoTek His Fab-Trap® Agarose

Validation Data Gallery

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  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).

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  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).

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  IP of GFP-His fusion protein from transfected HEK293T cells using His Fab-Trap Agarose (hfa) followed by a two-step elution using His-Peptide (hp). I: Input, F: Flow-through, E1: 1st elution, E2: 2nd elution, R: Residual.

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  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.

The ChromoTek His Fab-Trap® Agarose consists of an anti-His-tag Fab fragment, which is covalently bound to agarose beads. It can be used for the immunoprecipitation of proteins fused to the His-tag from cell extracts of various organisms.

Cat No. hfa

Reviews (1)

Specificity

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

Applications

IP, Co-IP

Type

Fab-fragment

Conjugate

Agarose

Synonyms

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

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Product Information

The ChromoTek His Fab-Trap® Agarose consists of an anti-His-tag Fab fragment, which is covalently bound to agarose beads. It can be used for the immunoprecipitation of proteins fused to the His-tag from cell extracts of various organisms.

• Description: Immunoprecipitation of His-tagged proteins with anti-His-tag Fab-fragment conjugated to agarose beads.;

  • 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.

• Applications: IP, Co-IP
• Specificity/Target: Binds 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 Capacity: 25 ug of recombinant His-tagged protein (~30kDa) per 25 uL bead slurry
• Conjugate: Agarose beads; ~90 um (cross-linked 4% agarose beads)
• Elution buffer: 2x SDS-sample buffer (Lämmli), 200 mM glycine pH 2.5 or 100 µM His-Peptide
• Wash Buffer Compatibility: 2 M NaCl, 5 mM DTT, 5 mM β-mercaptoethanol, 5 mM TCEP, 2% NP40, 2% Triton X-100, 0.1% SDS, 5 M Urea
• Type: Fab-fragment
• Class: Recombinant
• Host: Mouse
• Affinity (KD): 10 nM for a C-terminal His-tag and 220 nM for a N-terminal His-tag
• Compatibility with mass spectrometry: The His Fab-Trap® is optimized for on-bead digestion. For the application note, please click here:; On-bead digest protocol for mass spectrometry
• RRID: AB_3661838
• Storage Buffer: PBS pH 7.4. Preservative: 0.09% sodium azide
• Storage Condition: Shipped at ambient temperature. Upon receipt store at +4°C. Stable for one year. Do not freeze!
• Size: 25ul/reactions (eg：20rxns=500ul slurry)

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Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Talita (Verified Customer) (09-27-2025)

It was my first time performing the purification using beads, and unfortunately it did not work very well. The protein yield was very low, so I performed the concentration using a centrifuge device. Subsequently, the protein quantification and followed by a western blotting. Unfortunately, the analysis was not successful and I was not able to detect the his-tagged protein. However, I would give this product another try due to its short protocol.

• Applications: Protein purification
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Gram-positive bacteria (P. gingivalis), culture supernatant