Human HAND1 Antibody

R&D Systems | Catalog # AF3168

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Immunocytochemistry, Simple Western

Cited:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP), Western Blots

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
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Product Specifications

Immunogen

E. coli-derived recombinant human HAND1
Met1-Gln215
Accession # O96004

Specificity

Detects human HAND1 in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant human HAND2 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human HAND1 Antibody

Detection of Human HAND1 antibody by Western Blot.

Detection of Human HAND1 by Western Blot.

Western blot shows lysates of human heart tissue, human brain (cerebellum) tissue, and human placenta tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human HAND1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3168) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for HAND1 at approximately 22 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

HAND1 in SH‑SY5Y Human Cell Line.

HAND1 was detected in immersion fixed SH‑SY5Y human neuroblastoma cell line using Goat Anti-Human HAND1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3168) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of Human HAND1 by Simple WesternTM.

Left: Simple Western lane view shows lysates of human cerebellum, loaded at 1 mg/ml. A specific band was detected for HAND1 at approximately 26 kDa (as indicated) using both 10 µg/ml and 50 µg/ml of Goat Anti-Human HAND1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3168) followed by HRP-conjugated Donkey Anti-Goat Secondary Antibody (Catalog # 043-522-2). This experiment was conducted under reducing conditions and using the 2-40kDa separation system. Right: Simple Western electropherogram showing the same Goat Anti-Human HAND1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3168) tested at 10 µg/ml (blue line) and 50 µg/ml (green line) in the human cerebellum.
Detection of HAND1 by Immunohistochemistry

Detection of HAND1 by Immunohistochemistry

Directed differentiation of small ape iPSCs. (A) Schematic overview of direct neurosphere formation and the subsequent neuronal differentiation cultures. (B) Phase-contrast images of representative neurospheres (day 7) and neurons (day 21) derived from small ape iPSCs. (C) Immunofluorescence images of beta III tubulin in day 21 neurons. Nuclei were counterstained with DAPI. Scale bar: 100 μm. (D) Schematic design of limb bud mesoderm differentiation culture. (E) Immunofluorescence images of HAND1 in day 2 lateral plate mesoderm cells and PRRX1 in day 4 limb bud mesoderm cells. Nuclei were counterstained with DAPI. Scale bar: 100 μm. (F) Heatmap showing the expression of genes associated with pluripotency (POU5F1, NANOG), lateral plate mesoderm (HAND1, FOXF1, NKX2.5, ISL1), and limb bud mesoderm (PRRX1, MSX1, PITX1) in iPSCs, day 2 lateral plate mesoderm cells, and day 4 limb bud mesoderm cells determined by quantitative RT-PCR analysis. beta -ACTIN was used as an internal control. (G) Immunofluorescence images of SOX17 in day 2 definitive endoderm cells. Nuclei were counterstained with DAPI. Scale bar: 100 μm. (H) Heatmap showing the expression of genes associated with pluripotency (POU5F1, NANOG) and definitive endoderm (SOX17, FOXA2, GSC, CER1, HHEX, GATA6) in iPSCs and day 2 definitive endoderm cells determined by quantitative RT-PCR analysis. Image collected and cropped by CiteAb from the following open publication (https://www.frontiersin.org/articles/10.3389/fcell.2025.1536947/full), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of HAND1 by Immunocytochemistry/ Immunofluorescence

Detection of HAND1 by Immunocytochemistry/ Immunofluorescence

Pseudotime trajectories of TGF beta -inhibited naïve hPSCs recapitulates early trophectoderm specification in human embryos.(a) Diffusion maps of naïve hPSC 10X scRNA-seq data during the SB time-course, reporting diffusion pseudotime scores. (b) Diffusion map visualisation of naïve cells during the SB time-course reporting the relative expression of additional pluripotency markers KLF4 and DPPA5, TGF beta effector NODAL, and trophoblast markers CDX1, MARCKS, and GATA2. (c) Immunofluorescence microscopy for OCT4 (magenta), HAND1 (yellow), GATA3 (cyan) and DAPI (blue) in H9 NK2 naïve cells following 3 days of SB supplementation to t2iLGö medium. White arrowheads indicate cells that co-express low levels of OCT4 and HAND1 or GATA3. Scale bars: 50 µm. (d) Correlation heatmap between the pseudobulked datasets from all Louvain clusters (A/B/C/D/E) and the identified cell lineages from the cultured human pre-gastrulation embryos (Xiang et al., 2020). CTB: Cytotrophoblast; EVT: Extravillous trophoblast; STB: Syncytiotrophoblast; EPI: Epiblast; ICM: Inner Cell Mass; PrE: Primitive Endoderm, TE: Trophectoderm. (e) Heatmap reporting the expression in the Louvain clusters A, D, and E of the top 20 differentially expressed genes between EPI, ICM, and CTB cell lineages (based on data published by Xiang et al., 2020). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34463252), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of HAND1 by Flow Cytometry

Detection of HAND1 by Flow Cytometry

Generation of hPSC-LV-CMs using a two-step approach(A) Depiction of the standard WNT-ON/WNT-OFF protocol; modifications to this protocol (±) and critical feeding steps are indicated. CHIR, CHIR99021; BMP, BMP4; A, activin A; FGF, FGF2; LAA, L-ascorbic acid; IWR1, WNT pathway inhibitor; AGN, AGN193109, a pan-retinoic acid (RA) inhibitor; VitA, vitamin A.(B) Immunostaining micrographs of day 20 hPSC-CMs generated from mesoderm cells exposed to activin A, FGF2, and varying amounts of BMP4 and CHIR, demonstrating the cardiac corridor for making LV-like cardiomyocytes (red box). Scale bar: 100 μm.(C) Flow cytometry plot of day 20 hPSC-LV-CM cultures.(D and E) Day 20 hPSC-CMs generated in the presence or absence of RA signaling (±AGN, ±vitamin A, ±RA).(D) Immunostaining micrographs; scale bar: 100 μm.(E) qRT-PCR analysis.(F and G) Cardiomyocyte quantification in day 20 hPSC-CM cultures differentiated using the standard (Std) or LV protocols. Cells were co-stained with ACTN2, MYL2, and HAND1. DAPI was used to normalize the data.See also Figure S2. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37159667), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human HAND1 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed SH‑SY5Y human neuroblastoma cell line

Simple Western

10-50 µg/mL
Sample: Human cerebellum tissue

Western Blot

1 µg/mL
Sample: Human heart tissue, human brain (cerebellum) tissue, and human placenta tissue

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.<br/>*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: HAND1

HAND1 belongs to the HAND family of conserved basic helix-loop-helix transcription factors. It is expressed in the heart, ganglia, neural crest derivatives and extraembryonic tissues. HAND1 plays an essential role in heart development and is critical for the specification of extraembryonic tissues including trophoblasts. Human and mouse HAND1 share approximately 92% amino acid sequence homology.

Long Name

Heart And Neural Crest Derivatives Expressed 1

Alternate Names

eHand, Thing1

Entrez Gene IDs

9421 (Human); 15110 (Mouse); 59112 (Rat)

Gene Symbol

HAND1

UniProt

Additional HAND1 Products

Product Documents for Human HAND1 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human HAND1 Antibody

For research use only

Citations for Human HAND1 Antibody

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Protocols

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