Name: Anti-HAND2 antibody [EPR19451]
Brand: RabMAb
SKU: ab200040
Rating: 5 (2 reviews)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAND2 antibody [EPR19451] (ab200040)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19451] to HAND2
Suitable for: IHC-P, WB, ICC/IF, IP, Flow Cyt (Intra)
Reacts with: Mouse, Rat, Human

Carrier Free

Product name

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

Rabbit monoclonal [EPR19451] to HAND2
Host species

Rabbit
Tested applications

Suitable for: IHC-P, WB, ICC/IF, IP, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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 (intra): Neuro-2a cells. IP: SH-SY5Y whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19451
Isotype

IgG
Research areas

The Abpromise guarantee

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
IHC-P	(1)	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.
Flow Cyt (Intra)		1/400.

Notes
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.
Flow Cyt (Intra) 1/400.

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.
Target information above from: UniProt accession P61296 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 9464 Human
- Entrez Gene: 15111 Mouse
- Entrez Gene: 64637 Rat
- Omim: 602407 Human
- SwissProt: P61296 Human
- SwissProt: Q61039 Mouse
- SwissProt: P61295 Rat
- Unigene: 388245 Human
- Unigene: 430844 Mouse
- Unigene: 41057 Rat
see all
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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Western blot - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Western blot - Anti-HAND2 antibody [EPR19451] (ab200040)

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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Immunocytochemistry/ Immunofluorescence - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Immunocytochemistry/ Immunofluorescence - Anti-HAND2 antibody [EPR19451] (ab200040)

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 Cytometry (Intracellular) - Anti-HAND2 antibody [EPR19451] (ab200040)

Intracellular 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 Fluorr^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Anti-HAND2 antibody [EPR19451] (ab200040)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

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Datasheet download

Download

Publishing research using ab200040? Please let us know so that we can cite the reference in this datasheet.

ab200040 has been referenced in 7 publications.

Zhang D et al. Cysteine dioxygenase and taurine are essential for embryo implantation by involving in E2-ERα and P4-PR signaling in mouse. J Anim Sci Biotechnol 14:6 (2023). PubMed: 36604722
Manivannan S et al. Single-cell transcriptomic profiling unveils dysregulation of cardiac progenitor cells and cardiomyocytes in a mouse model of maternal hyperglycemia. Commun Biol 5:820 (2022). PubMed: 35970860
Liu Z et al. Loss of CASZ1 tumor suppressor linked to oncogenic subversion of neuroblastoma core regulatory circuitry. Cell Death Dis 13:871 (2022). PubMed: 36243768
Li K et al. Loss of basigin expression in uterine cells leads to subfertility in female mice†. Biol Reprod 105:859-875 (2021). PubMed: 34106247
Lewis-Israeli YR et al. Generating Self-Assembling Human Heart Organoids Derived from Pluripotent Stem Cells. J Vis Exp N/A:N/A (2021). PubMed: 34605811
Xia M et al. HAND2-mediated epithelial maintenance and integrity in cardiac outflow tract morphogenesis. Development N/A:N/A (2019). PubMed: 31201155
Han X et al. The lncRNA Hand2os1/Uph locus orchestrates heart development through regulation of precise expression of Hand2. Development 146:N/A (2019). PubMed: 31273086

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1-2 of 2 Abreviews

Application

Flow Cytometry

Sample

Human Cell (Human induced pluripotent stem cells)

Permeabilization

Yes - Triton-x100 0.5% in PBS

Gating Strategy

whole cell population

Specification

Human induced pluripotent stem cells

Fixation

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 10 2022

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

Specification

placenta

Blocking step

Serum as blocking agent for 20 minute(s) · Concentration: 2% · Temperature: 23°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Sep 06 2018

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Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Developmental stage

Cellular localization

Database links

Alternative names

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Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (7)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HAND2 antibody [EPR19451]