HHEX Antibody (2018B)

R&D Systems | Catalog # MAB83771

Recombinant Monoclonal Antibody.
R&D Systems
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Xenograft

Applications

Validated:

Western Blot, Flow Cytometry, Immunocytochemistry

Cited:

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

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2018B
View all HHEX Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human HHEX
Thr111-Gly270
Accession # Q03014

Specificity

Detects human HHEX in direct ELISAs and Western blots.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for HHEX Antibody (2018B)

Detection of Human, Mouse, and Rat HHEX antibody by Western Blot.

Detection of Human, Mouse, and Rat HHEX by Western Blot.

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, BaF3 mouse pro-B cell line, and H4-II-E-C3 rat hepatoma cell line. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human/Mouse/Rat HHEX Monoclonal Antibody (Catalog # MAB83771) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for HHEX at approximately 37 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
HHEX antibody in K562 Human Cell Line by Immunocytochemistry (ICC).

HHEX in K562 Human Cell Line.

HHEX was detected in immersion fixed K562 human chronic myelogenous leukemia cell line using Rabbit Anti-Human/Mouse/Rat HHEX Polyclonal Antibody (Catalog # MAB83771) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of HHEX antibody in A549 Human Cell Line antibody by Flow Cytometry.

Detection of HHEX in A549 Human Cell Line by Flow Cytometry.

A549 human lung carcinoma cell line was stained with Rabbit Anti-Human/Mouse/Rat HHEX Monoclonal Antibody (Catalog # MAB83771, filled histogram) or isotype control antibody (Catalog # AB-105-C, open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
HHEX antibody in iBJ6 iPS Cell Line by Immunocytochemistry (ICC).

HHEX in iBJ6 iPS Cell Line.

HHEX was detected in immersion fixed iBJ6 iPS cell line differentiated into hepatocytes using Rabbit Anti-Human/Mouse/Rat HHEX Polyclonal Antibody (Catalog # MAB83771) at 2 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of bl2vlcbkhad3mwbg8e4s.jpg by Immunocytochemistry/ Immunofluorescence

Detection of bl2vlcbkhad3mwbg8e4s.jpg by Immunocytochemistry/ Immunofluorescence

Suppressive effect of HHEX on the neural feature of SH-SY5Y cells. (A) Phenotypic alteration of SH-SY5Y cells in response to enforced expression of HHEX after 4 days of virus infection. Cells infected with lentivirus derived from empty vector were used as control (Vector). (B) IF detection of expression of neuronal and neural stemness proteins in cells with enforced HHEX expression. Cell nuclei were counterstained with DAPI. (C) IB detection of expression of a series of proteins in response to HHEX expression. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34595169), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of inkpsgf4a3xeawdwz7f9.jpg by Western Blot

Detection of inkpsgf4a3xeawdwz7f9.jpg by Western Blot

Suppressive effect of HHEX on the neural feature of SH-SY5Y cells. (A) Phenotypic alteration of SH-SY5Y cells in response to enforced expression of HHEX after 4 days of virus infection. Cells infected with lentivirus derived from empty vector were used as control (Vector). (B) IF detection of expression of neuronal and neural stemness proteins in cells with enforced HHEX expression. Cell nuclei were counterstained with DAPI. (C) IB detection of expression of a series of proteins in response to HHEX expression. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34595169), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of HHEX by Western Blot

Detection of HHEX by Western Blot

HHEX or Foxa3 suppresses malignant features or tumorigenicity in U118MG and NE-4C cells. (A–D) Effect of enforced expression of HHEX or Foxa3 on U118MG cells. (A) Phenotypic change in U118MG cells in response to enforced expression of HHEX or Foxa3. IB detection of a series of cancer-promoting factors and neuronal markers in cells with and without (Vector) enforced expression of HHEX (B) or Foxa3 (C). IB was performed with whole cell lysate. b-ACT was detected as a loading control. (D) Effect of HHEX or Foxa3 expression on the capability of cell migration and invasion. (E–M) Effect of enforced expression of HHEX or Foxa3 on NE-4C cells. (E) The influence of HHEX or Foxa3 expression on neurosphere formation in NSC specific serum-free medium in the time period as indicated. RT-qPCR detection of changes in transcription of genes representing neural stemness, neuronal differentiation, and genes involved in the basic cellular physiological machineries in control cells (Vector) and cells with HHEX (F) or Foxa3 (G) expression. Significance in change of transcription level was calculated for experiments in triplicate using unpaired Student’s t-test. Data are shown as mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001. NS, not significant. Effect of expression of Foxa3 (H–J) or HHEX (K–M) on tumorigenicity of NE-4C, as assayed with xenograft tumor formation (H,K) in immunodeficient nude mice. Significance of difference in tumor volumes (I,L) between two groups of mice was calculated using two-way ANOVA-Bonferroni/Dunn test. Significance of difference in tumor weight (J,M) was calculated using unpaired Student’s t-test. Data are shown as mean ± SEM. *p < 0.05 and ***p < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34595169), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of HHEX by Western Blot

Detection of HHEX by Western Blot

HHEX, Foxa3, or their combination suppresses neural stemness and tumorigenicity in HCT116 cells. (A) Morphological alteration in HCT116 cells in normal culture medium and change in ability of neurosphere-like formation in NSC-specific serum-free medium in response to enforced expression of HHEX, Foxa3, or their combination. (B) IB detection of protein expression alteration in response to enforced expression of HHEX, Foxa3, and their combination. (C,D) Effect of expression HHEX, Foxa3, or their combination on invasion, migration, and colony formation in soft agar (C). Significance in difference of colony formation was calculated based on experiments in triplicate using unpaired Student’s t-test (D). Colonies larger than 100 μm in diameter were counted. Data are shown as mean ± SEM. *p < 0.05 and **p < 0.01. (E–G) Xenograft tumor formation (E) by control HCT116 cells (Vector) and cells with enforced expression of HHEX, Foxa3, and both. Significance of difference in tumor weight between two groups of mice was calculated using unpaired Student’s t-test (F), and significance of difference in tumor volume was calculated using two-way ANOVA-Bonferroni/Dunn test (G). Data are shown as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34595169), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of HHEX by Western Blot

Detection of HHEX by Western Blot

Detection of expression effect of HHEX and Foxa3 in cells and colon tissues, and the effect on colon tumor formation in AOM/DSS model of colitis-associated cancer. (A) Detection of expression effect of AAV-HHEX and AAV-Foxa3 constructs in HEK293T cells. Nuclei were counterstained with DAPI. (B) Schematic illustration of the strategy for generation of AOM/DSS model of coloretal cancer and time points for AAV administration. (C) Body weight change of the wild type mice and mice with AOM/DSS treatment. Significance in body weight between two groups was calculated using two-way ANOVA-Bonferroni/Dunn test. Data are shown as mean ± SEM. NS, not significant. (D) Detection of expression effect of the infected AAV virus in colon tissue in different groups via observing GFP in cryosections. (E) IB detection of HHEX and Foxa3 expression effect in colon tissues. (F) Colons harvested from three groups of mice, as indicated. (G) Colonic weight/length ratio in three groups of mice as indicated. (H) Numbers of tumors observed in three groups of mice as indicated. In (G,H), significance was calculated using unpaired Student’s t-test. Data are shown as mean ± SEM. **p < 0.01, ***p < 0.001, and ****p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34595169), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for HHEX Antibody (2018B)

Application
Recommended Usage

Flow Cytometry

0.25 µg/106 cells
Sample: A549 human lung carcinoma cell line fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012)

Immunocytochemistry

2-25 µg/mL
Sample: Immersion fixed K562 human chronic myelogenous leukemia cell line and immersion fixed iBJ6 iPS cell line differentiated into hepatocytes

Western Blot

1 µg/mL
Sample: HepG2 human hepatocellular carcinoma cell line, BaF3 mouse pro-B cell line, and H4‑II‑E‑C3 rat hepatoma cell line

Reviewed Applications

Read 2 reviews rated 5 using MAB83771 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *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: HHEX

Hematopoietically-expressed homeobox protein (HHEX), also known as HEX, PRH and PRHX, is a 35-40 kDa member of the Homeobox family of transcription factors.  Family members are distinguished by an evolutionarily conserved DNA-binding homeodomain of 60 amino acids (aa), which for HHEX spans aa 137-196.  Human HHEX was initially isolated from hematopoietic tissue, and is present in several hematopoietic progenitors, where its expression is down-regulated during terminal cell differentiation.  HHEX is also expressed in the anterior visceral endoderm during early mouse development, and in some adult tissues of endodermal origin, including liver, lung and thyroid.  HHEX knockout in mice is embryonic lethal, with impaired forebrain, liver and thyroid development.  Human HHEX is 270 aa in length, and over aa 111-270, shares 93% and 95% identity with mouse and rat HHEX, respectively.

Long Name

Hematopoietically Expressed Homeobox

Alternate Names

Hex, HMPH, HOX11L-PEN, PRH, PRHX

Entrez Gene IDs

3087 (Human); 15242 (Mouse); 79237 (Rat)

Gene Symbol

HHEX

UniProt

Q03014

Additional HHEX Products

Product Documents for HHEX Antibody (2018B)

Certificate of Analysis

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Product Specific Notices for HHEX Antibody (2018B)

For research use only

Citations for HHEX Antibody (2018B)

Customer Reviews for HHEX Antibody (2018B) (2)

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  • Human/Mouse/Rat HHEX Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: HT-29 human colon adenocarcinoma cell line
    Species: Human
    Verified Customer | Posted 07/09/2022
    HHEX Antibody (2018B) MAB83771
  • Human/Mouse/Rat HHEX Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Osteoblasts
    Species: Mouse
    Verified Customer | Posted 11/03/2021
    HHEX Antibody (2018B) MAB83771

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