Anti-HA tag Rabbit mAb - ChIP, CUT&RUN and CUT&Tag Grade

Catalog number :AT1747

Overview

Tested applications: WB : 1/1000-1/5000.

IHC : 1/200-1/500.

ICC/IF : 1/100-1/500.

IP : 1/50-1/200.

ChIP : 1/50-1/100.
CUT&RUN and CUT&Tag

Optimal dilutions/concentrations should be determined by the end user.

Specificity: HA Antibody Recognizes HA-tagged proteins overexpressed in cells, including both amino- and carboxy-termini of targeted proteins in transfected mammalian cells.

Properties

Applications

WB Image: All lanes: Anti-HA tag antibody- ChIP grade- (AT1747) at 1/5000 dilution

Lane 1: HEK293 human embryonic kidney cell line transfected with N-terminal HA-tagged MICA

Lane 2: HEK293 human embryonic kidney cell line transfected with C-terminal HA-tagged TRANCE

Lane 3: HEK293 human embryonic kidney cell line transfected with C-terminal HA-tagged LRRN5

Recommended secondary antibody: Goat anti rabbit IgG-HRP (Engibody, Catalog number:AT0097)

Recommended ECL: ECL Pico-Detect™ Western Chemiluminescent HRP Substrate (Engibody, Catalog number: IF6747)

ICC/IF Image: Immunocytochemistry:

in immersion fixed NS0 mouse myeloma cell line transfected with HA-tagged WISP-2 using Rabbit Anti-HA Monoclonal Antibody – ChIP Grade

Recommended secondary antibody: Goat Anti-Rabbit IgG(H+L) (IFluor™ 594) (Engibody, Catalog number:AT1553)

ChIP Images: Xenopus laevis oocytes were injected with mRNA for HA-tagged human BORIS. Chromatin was prepared according to the ChIP protocol. Oocytes were fixed with formaldehyde for 10min.

The ChIP was performed with 25µg of chromatin, 20µl of Protein A/G agarose beads, and 5µg of anti-HA antibody, light blue) or, 5µg of anti-Boris antibody, dark blue). A non-specific antibody was used as a control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach).