Overview

  • Product name

    Anti-HAND2 antibody [EPR19451]
    See all HAND2 primary antibodies
  • Description

    Rabbit monoclonal [EPR19451] to HAND2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, IHC-P, WB, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse HAND2 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q61039

  • Positive control

    • WB: HEK-293T transfected with mouse HAND2 expression vector containing a myc tag, whole cell lysate; Neuro-2a and SH-SY5Y whole cell lysates. IHC-P: Mouse E14.5 embryo and E14.5 embryonic adrenal gland tissues; Rat E14.5 embryo and E14.5 embryonic gut tissues. ICC/IF: Neuro-2a and SH-SY5Y cells. Flow Cyt: Neuro-2a cells. IP: SH-SY5Y whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab200040 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/400.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

The human species recommendation is based on the WB results. We do not guarantee IHC-P for human species.

WB 1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 24 kDa).
ICC/IF 1/100.
IP 1/30.

Target

  • Function

    Essential for cardiac morphogenesis, particularly for the formation of the right ventricle and of the aortic arch arteries. Required for vascular development and regulation of angiogenesis, possibly through a VEGF signaling pathway. Plays also an important role in limb development, particularly in the establishment of anterior-posterior polarization, acting as an upstream regulator of sonic hedgehog (SHH) induction in the limb bud. Is involved in the development of branchial arches, which give rise to unique structures in the head and neck. Binds DNA on E-box consensus sequence 5'-CANNTG-3'.
  • Tissue specificity

    Heart.
  • Sequence similarities

    Contains 1 basic helix-loop-helix (bHLH) domain.
  • Developmental stage

    Expressed in the fetal heart.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • AI225906 antibody
    • AI661148 antibody
    • autonomic nervous system and neural crest derivatives-expressed protein 2 antibody
    • Basic helix loop helix transcription factor HAND2 antibody
    • bHLHa26 antibody
    • Class A basic helix-loop-helix protein 26 antibody
    • Deciduum antibody
    • Deciduum heart autonomic nervous system and neural crest derivatives expressed protein 2 antibody
    • DHAND 2 antibody
    • dHAND antibody
    • DHAND2 antibody
    • Ehand 2 antibody
    • Ehand2 antibody
    • FLJ16260 antibody
    • HAND 2 antibody
    • Hand2 antibody
    • HAND2_HUMAN antibody
    • Heart and neural crest derivatives expressed 2 antibody
    • Heart and neural crest derivatives expressed protein 2 antibody
    • heart and neural crest derivatives expressed transcript 2 antibody
    • heart antibody
    • Heart- and neural crest derivatives-expressed protein 2 antibody
    • Hed antibody
    • Highly similar to dHAND M musculus antibody
    • MGC125303 antibody
    • MGC125304 antibody
    • Th 2 antibody
    • Th2 antibody
    • Thing 2 antibody
    • Thing2 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded mouse E14.5 embryo tissue labeling HAND2 with ab200040 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in multiple tissues of E14.5 mouse embryo (PMID: 25248475). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • All lanes : Anti-HAND2 antibody [EPR19451] (ab200040) at 1/1000 dilution

    Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with an empty vector (vector control) containing a myc tag, whole cell lysate
    Lane 2 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with mouse HAND2 expression vector containing a myc tag, whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 24 kDa
    Observed band size: 26 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The plasmid was kindly provided by Dr. Zhongzhou Yang from Nanjing University.

  • All lanes : Anti-HAND2 antibody [EPR19451] (ab200040) at 1/1000 dilution

    Lane 1 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysate
    Lane 2 : SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 24 kDa
    Observed band size: 25 kDa why is the actual band size different from the predicted?



     

    Exposure times: Lane 1: 3 minutes; Lane 2: 10 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular mass observed is consistent with what has been described in the literature (PMID: 26166202); the upper band could be ubiquitinated Hand2 (PMID 21170031).

  • Immunohistochemical analysis of paraffin-embedded mouse E14.5 embryonic adrenal gland tissue labeling HAND2 with ab200040 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in mouse embryonic (E14.5) ganglion and adrenal gland (PMID: 25248475). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded rat E14.5 embryonic tissue labeling HAND2 with ab200040 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in multiple rat embryonic (E14.5) tissues (PMID: 25248475). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded rat E14.5 embryonic gut tissue labeling HAND2 with ab200040 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in rat E14.5 embryonic gut (PMID: 25248475). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling HAND2 with ab200040 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Nuclear staining in Neuro-2a cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (human neuroblastoma cell line from bone marrow) cells labeling HAND2 with ab200040 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Nuclear staining in SH-SY5Y cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Neuro-2a (mouse neuroblastoma cell line) cell line labeling HAND2 with ab200040 at 1/400 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

  • HAND2 was immunoprecipitated from 0.35 mg of SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate with ab200040 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab200040 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: SH-SY5Y whole cell lysate 10 µg (Input). 

    Lane 2: ab200040 IP in SH-SY5Y whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200040 in SH-SY5Y whole cell lysate.

    Exposure time: Less than 1 second.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

References

This product has been referenced in:

  • Xia M  et al. HAND2-mediated epithelial maintenance and integrity in cardiac outflow tract morphogenesis. Development N/A:N/A (2019). Read more (PubMed: 31201155) »
See 1 Publication for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rhesus monkey Tissue sections (placenta)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate ph6.0
Permeabilization
No
Specification
placenta
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 2% · Temperature: 23°C
Fixative
Paraformaldehyde

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Submitted Sep 06 2018

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