Human PLZF Antibody (6318100)

R&D Systems | Catalog # MAB2944

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP)

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 6318100
View all PLZF Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human PLZF
Met1-Gln254
Accession # Q05516

Specificity

Detects human PLZF in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant human (rh) ZBTB38, rhZNF24, rhZNF143, rhZNF206, rhZNF281, or rhZNF423 is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human PLZF Antibody (6318100)

Detection of Human PLZF antibody by Western Blot.

Detection of Human PLZF by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, HEL 92.1.7 human erythroleukemic cell line, LNCaP human prostate cancer cell line, and 293T human embryonic kidney cell line. PVDF Membrane was probed with 1 µg/mL of Human PLZF Monoclonal Antibody (Catalog # MAB2944) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for PLZF at approximately 85 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
PLZF antibody in HL-60 Human Cell Line by Immunocytochemistry (ICC).

PLZF in HL‑60 Human Cell Line.

PLZF was detected in immersion fixed HL-60 human acute promyelocytic leukemia cell line using Human PLZF Monoclonal Antibody (Catalog # MAB2944) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel; Catalog # NL007) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of antibody in HL-60 Human Cell Line antibody by Flow Cytometry.

Detection of in HL‑60 Human Cell Line by Flow Cytometry.

HL-60 human acute promyelocytic leukemia cell line was stained with Human PLZF Monoclonal Antibody (Catalog # MAB2944, filled histogram) or isotype control antibody (Catalog # MAB003, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
Detection of PLZF by Western Blot

Detection of PLZF by Western Blot

Spermatogenic differentiation of YAP double knockdown hESCs (YAP-DKD cells) (A). Western blot shows the level of YAP relative to beta -ACTIN in YAP-DKD cells compared to that of YAP-KD cells and wild-type hESCs (WT). ****P < 0.0001. (B) Graphs show the expression levels of YAP target genes, CTGF and CYR61, in YAP-DKD cells compared to those of YAP-KD cells and WT. Data are presented as mean ± SD of 3 experiments. **P < 0.01, ***P < 0.001. (C) Western blot shows YAP, PLZF, VASA and acrosin protein levels relative to beta -ACTIN in WT, YAP-KD, and YAP-DKD cells during their spermatogenic differentiation. (D) Graphs show the level of PLZF protein relative to beta -ACTIN in YAP-DKD cells during their spermatogenic differentiation compared to YAP-KD cells and WT. (E) Graphs show the expression levels of spermatogenic cell markers (VASA, PLZF, GFRA1, GPR125 and ACR) during spermatogenic differentiation of YAP-DKD, compared to those of YAP-KD cells and WT. Data are presented as means ± SD of 3 experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (F) Representative micrographs show the morphology of YAP-DKD cells during their spermatogenic differentiation. White dots indicate areas of dead cells. Scale bar: 200 μm for the upper panel and 100 μm for the lower panel. (G) Western blot shows the level of the apoptotic marker cleaved Caspase3 relative to beta -ACTIN in WT and YAP-DKD cells during their spermatogenic differentiation. The uncropped membranes associated with this figure are provided in Supplementary Fig. S10. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41598-024-66852-x), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of PLZF by Immunohistochemistry

Detection of PLZF by Immunohistochemistry

G3BP2 deficiency-induced germ cell apoptosis.(A) Cell apoptosis detected by TUNEL assay from WT, SKO, and DKO adult mice. Apoptotic cells are marked red and nuclei were stained DAPI, scale bar = 100 μm. (B) Tunel positive cells in 100 tubules at WT, SKO, and DKO adult mice. Data are presented as mean ± S D, n = 5. ***, p < 0.001 by student’s t-test. (C) Immunodetection of PLZF in testis sections from adult WT, SKO, and DKO mice. Scale bar: 50 μm. Quantification of PLZF-positive cells in seminiferous tubule from WT, SKO, and DKO mice. A total of 20 tubules were counted. (D) Heat stress adult testis sections stained for TIA1, a marker of stress granules. Magnified images are shown at the bottom. The condensed stress granules are marked by white arrow. Scale bar: 20 μm. Quantification of TIA1-positive cells in seminiferous tubule from WT, SKO, and DKO mice. A total of 20 tubules were counted. NS, no significant. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35782098), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of PLZF by Immunocytochemistry/ Immunofluorescence

Detection of PLZF by Immunocytochemistry/ Immunofluorescence

Modulation of YAP expression during the various stages of spermatogenic cell differentiation of wild-type hESCs by using DH and LPA. (A) Diagrams show the timeline of DH and LPA supplementation during spermatogenic differentiation of wild-type hESCs. (B) Graphs show the total number of differentiated hESCs and the number of PLZF+ hSSCs derived from various conditions described in (A) at the end of the culture. Data are presented as mean ± SD of 3 experiments. **P < 0.01, ***P < 0.001, ****P < 0.0001 and ### P < 0.001 indicated the statistically significant of spermatogonia number from DH treated cells between each group. (C) Representative immunofluorescence micrographs show the level of PLZF expression in DH-ET and LPA-ET cells compared to that of their untreated counterpart. Scale bar: 200 μm. (D) Western blot shows the expression of YAP, PLZF, Acrosin, and beta -ACTIN proteins in DH-ET cells during their spermatogenic differentiation compared to those of DH-AT cells, untreated control cells, and non-differentiated hESCs. Red numbers indicate the expression level of each sample relative to beta -ACTIN. (E) Analysis of DNA content by flow cytometry shows the percentage of haploid spermatid-like cells derived from DH-ET cells at the end of their spermatogenic differentiation compared to that of LPA-ET and untreated cells. (F) The graph shows the percentage of haploid spermatid-like cells derived from DH-ET cells at the end of their spermatogenic differentiation compared to that of LPA-ET and untreated cells. Data are presented as mean ± SD of 3 experiments. **P < 0.01, ***P < 0.001. The uncropped membranes associated with Fig. 5D are provided in Supplementary Fig. S11. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41598-024-66852-x), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of PLZF by Western Blot

Detection of PLZF by Western Blot

Modulation of YAP expression during the various stages of spermatogenic cell differentiation of wild-type hESCs by using DH and LPA. (A) Diagrams show the timeline of DH and LPA supplementation during spermatogenic differentiation of wild-type hESCs. (B) Graphs show the total number of differentiated hESCs and the number of PLZF+ hSSCs derived from various conditions described in (A) at the end of the culture. Data are presented as mean ± SD of 3 experiments. **P < 0.01, ***P < 0.001, ****P < 0.0001 and ### P < 0.001 indicated the statistically significant of spermatogonia number from DH treated cells between each group. (C) Representative immunofluorescence micrographs show the level of PLZF expression in DH-ET and LPA-ET cells compared to that of their untreated counterpart. Scale bar: 200 μm. (D) Western blot shows the expression of YAP, PLZF, Acrosin, and beta -ACTIN proteins in DH-ET cells during their spermatogenic differentiation compared to those of DH-AT cells, untreated control cells, and non-differentiated hESCs. Red numbers indicate the expression level of each sample relative to beta -ACTIN. (E) Analysis of DNA content by flow cytometry shows the percentage of haploid spermatid-like cells derived from DH-ET cells at the end of their spermatogenic differentiation compared to that of LPA-ET and untreated cells. (F) The graph shows the percentage of haploid spermatid-like cells derived from DH-ET cells at the end of their spermatogenic differentiation compared to that of LPA-ET and untreated cells. Data are presented as mean ± SD of 3 experiments. **P < 0.01, ***P < 0.001. The uncropped membranes associated with Fig. 5D are provided in Supplementary Fig. S11. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41598-024-66852-x), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human PLZF Antibody (6318100)

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed HL-60 human acute promyelocytic leukemia cell line

Intracellular Staining by Flow Cytometry

2.5 µg/106 cells
Sample: HL‑60 human acute promyelocytic leukemia cell line, fixed with paraformaldehyde and permeabilized with saponin

Western Blot

1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line, HEL 92.1.7 human erythroleukemic cell line, LNCaP human prostate cancer cell line, and 293T human embryonic kidney cell line

Reviewed Applications

Read 1 review rated 5 using MAB2944 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: PLZF

Human PLZF is a 74 kDa nuclear protein that belongs to the POK family of transcriptional repressors. It is a 673 amino acid protein that contains an N-terminal BTB domain, followed by an acidic domain, a proline-rich region and a C-terminal zinc-finger domain. PLZF forms homodimers with RAR alpha and LAZ3 with its zinc-finger region. Alternate splice forms exist which are tissue-specific and show a deletion of either the BTB domain, the acidic region, or the proline-rich region. Human PLZF shares 96%, 97%, 96%, and 96% amino acid identity with rat, mouse, canine, and bovine PLZF, respectively.

Long Name

Promyelocytic Leukemia Zinc Finger

Alternate Names

ZBTB16, ZNF145

Entrez Gene IDs

7704 (Human); 235320 (Mouse)

Gene Symbol

ZBTB16

UniProt

Q05516

Additional PLZF Products

Product Documents for Human PLZF Antibody (6318100)

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Product Specific Notices for Human PLZF Antibody (6318100)

For research use only

Citations for Human PLZF Antibody (6318100)

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  • Human PLZF Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: human neural stem cells
    Species: Human
    Verified Customer | Posted 08/31/2020
    Human neural stem cells, MAB2944 used at a dilution of 1/200
    Human PLZF Antibody (6318100) MAB2944

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