TRIM8 Antibody - BSA Free

Immunocytochemistry/Immunofluorescence: TRIM8 Antibody [NBP1-89776] - Immunofluorescent staining of human cell line U-251 MG shows localization to nucleoplasm & vesicles.

Staining of human skeletal muscle shows strong cytoplasmic positivity in myocytes.

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Flow Cytometry: TRIM8 Antibody [NBP1-89776] - Knockdown of TRIM8 promotes differentiation & impairs GBM stemness & self‐renewal capacity. (A) Western blot analysis shows reduced CD133, NESTIN, SOX2, & c‐MYC, & upregulated GFAP in GBM neurosphere cells following TRIM8 knockdown by TRIM8 shRNA1 or TRIM8 shRNA2. NT shRNA = nontargeting control shRNA. Western blot also shows upregulated PIAS3 & reduced phosphorylated STAT3 (Tyr705) following shRNA knockdown of TRIM8. (B) ICC shows reduced expression of TRIM8 (red), NESTIN (white), & SOX2 (red) in TRIM8 shRNA1‐knockdown cells compared to NT shRNA cells. Nuclei were counterstained with DAPI (blue). Scale bar = 10 μm. (C) Flow cytometric analysis & statistical analysis of SOX2 expression in TRIM8‐overexpressing neurosphere cells. SOX2 was tagged by APC (blue: 676 nm) with TRIM8 tagged by PE (red: 555 nm). ***P < 0.001, ****P < 0.0001. Statistics: Data are means ± SD (n = 3), by nonparametric t‐test. (D,E) TRIM8 knockdown impairs neurosphere formation. Representative images of neurosphere expressing NT shRNA & TRIM8 shRNA1 at specific time points are shown. Scale bar = 30 μm. Statistical analysis of average sphere size: by one‐way analysis of variance (ANOVA). Data are means ± standard deviation (SD) (n = 6). **P < 0.01, ***P < 0.001, ****P < 0.0001. (F) Western blot analysis shows reduced TRIM8 & activated STAT3 (Tyr705), with increased GFAP expression following differentiation induced by serum. (G) ICC reveals reduced TRIM8 (red) & increased GFAP (white) after four days of differentiation induced by serum. Nuclei were counterstained with DAPI (blue). Scale bar = 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28100038), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Flow Cytometry: TRIM8 Antibody [NBP1-89776] - Ectopic expression of TRIM8 enhances stemness & self‐renewal of GBM‐derived neurospheres. (A) Western blot analysis shows upregulation of CD133, NESTIN, SOX2, & c‐MYC following ectopic expression of TRIM8‐GFP fusion protein. WB also shows reduced PIAS3 & increased phosphorylated STAT3 (Tyr705) & total STAT3 following TRIM8 overexpression. (B) ICC staining of TRIM8 (red) with stem cell markers NESTIN (white), & SOX2 (red) in neurosphere cells overexpressing TRIM8. Nuclei were counterstained with DAPI (blue). Scale bar = 10 μm. (C) Flow cytometric analysis & statistical analysis showing CD133 & SOX2 upregulation in neurosphere cells with TRIM8 overexpression. CD133 was tagged by PE (red: 555 nm) with TRIM8 tagged by APC (blue: 676 nm). SOX2 was tagged by APC (blue: 676 nm) with TRIM8 tagged by PE (red: 555 nm). ***P < 0.001, ****P < 0.0001. Statistics: Data are means ± SD (n = 3), by nonparametric t‐test. (D,E) Effects of TRIM8 overexpression on neurosphere formation. Representative images of neurospheres expressing GFP vector or TRIM8‐GFP vector at specific points. Scale bar = 30 μm. Statistical analysis of average sphere size: by one‐way analysis of variance (ANOVA). Data are means ± standard deviation (SD) (n = 6). **P < 0.01, ***P < 0.001, ****P < 0.0001. (F) Extreme limiting dilution assay (ELDA) shows that TRIM8 overexpression reduces the cell dose required for colony formation, supporting enhanced stem cell properties. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28100038), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

GERP-Antibody-Immunocytochemistry-Immunofluorescence-NBP1-89776-img0007.jpg

Immunocytochemistry/ Immunofluorescence: TRIM8 Antibody [NBP1-89776] - TRIM8 is expressed in GBM samples & neurosphere cells. (A–C) Gene expression correlation of TRIM8 with STAT3,SOX2, & NESTIN using U133 mRNA data from TCGA. TRIM8 vs STAT3: Pearson's correlation coefficient: r = 0.36, P < 0.00001. TRIM8 vs SOX2: Pearson's correlation coefficient: r = 0.22, P < 0.00001. TRIM8 vs NESTIN: Pearson's correlation coefficient: r = 0.22, P < 0.00001. (D) Schematic representation of TRIM8 protein with RING, B‐box, & coiled‐coil domains. (E) TRIM8 copy number variation among 577 GBM samples in TCGA dataset. TRIM8 shows hemizygous deletion in 88.04% of cases. (F) Western blot analysis of TRIM8 protein among six normal human brain tissues (N1–N6) & five GBMs (G1–G5). (G) Immunohistochemical (IHC) staining of TRIM8 in normal brain & GBM tissues. TRIM8 (+) cells (indicated by arrow) are predominantly neurons in the normal brain & show cytoplasmic staining. TRIM8 expression in GBM is predominant in nuclei of tumor cells with modest cytoplasmic staining. Sections were counterstained with hematoxylin. Scale bar = 10 μm. (H) Reverse transcription PCR analysis & western blot analysis of TRIM8 among two normal cell lines & six patient‐derived GBM neurosphere cells. NHNP = normal human neural progenitor cell. (I) Immunocytochemical (ICC) staining of TRIM8 in neurosphere cells showing nuclear expression of TRIM8. TRIM8 was labeled in GFP, & nuclei were counterstained with DAPI (blue). Scale bar = 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28100038), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: TRIM8 Antibody [NBP1-89776] - Ectopic expression of TRIM8 enhances stemness & self‐renewal of GBM‐derived neurospheres. (A) Western blot analysis shows upregulation of CD133, NESTIN, SOX2, & c‐MYC following ectopic expression of TRIM8‐GFP fusion protein. WB also shows reduced PIAS3 & increased phosphorylated STAT3 (Tyr705) & total STAT3 following TRIM8 overexpression. (B) ICC staining of TRIM8 (red) with stem cell markers NESTIN (white), & SOX2 (red) in neurosphere cells overexpressing TRIM8. Nuclei were counterstained with DAPI (blue). Scale bar = 10 μm. (C) Flow cytometric analysis & statistical analysis showing CD133 & SOX2 upregulation in neurosphere cells with TRIM8 overexpression. CD133 was tagged by PE (red: 555 nm) with TRIM8 tagged by APC (blue: 676 nm). SOX2 was tagged by APC (blue: 676 nm) with TRIM8 tagged by PE (red: 555 nm). ***P < 0.001, ****P < 0.0001. Statistics: Data are means ± SD (n = 3), by nonparametric t‐test. (D,E) Effects of TRIM8 overexpression on neurosphere formation. Representative images of neurospheres expressing GFP vector or TRIM8‐GFP vector at specific points. Scale bar = 30 μm. Statistical analysis of average sphere size: by one‐way analysis of variance (ANOVA). Data are means ± standard deviation (SD) (n = 6). **P < 0.01, ***P < 0.001, ****P < 0.0001. (F) Extreme limiting dilution assay (ELDA) shows that TRIM8 overexpression reduces the cell dose required for colony formation, supporting enhanced stem cell properties. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28100038), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

TRIM8 is expressed in GBM samples and neurosphere cells. (A–C) Gene expression correlation of TRIM8 with STAT3,SOX2, and NESTIN using U133 mRNA data from TCGA. TRIM8 vs STAT3: Pearson's correlation coefficient: r = 0.36, P < 0.00001. TRIM8 vs SOX2: Pearson's correlation coefficient: r = 0.22, P < 0.00001. TRIM8 vs NESTIN: Pearson's correlation coefficient: r = 0.22, P < 0.00001. (D) Schematic representation of TRIM8 protein with RING, B‐box, and coiled‐coil domains. (E) TRIM8 copy number variation among 577 GBM samples in TCGA dataset. TRIM8 shows hemizygous deletion in 88.04% of cases. (F) Western blot analysis of TRIM8 protein among six normal human brain tissues (N1–N6) and five GBMs (G1–G5). (G) Immunohistochemical (IHC) staining of TRIM8 in normal brain and GBM tissues. TRIM8 (+) cells (indicated by arrow) are predominantly neurons in the normal brain and show cytoplasmic staining. TRIM8 expression in GBM is predominant in nuclei of tumor cells with modest cytoplasmic staining. Sections were counterstained with hematoxylin. Scale bar = 10 μm. (H) Reverse transcription PCR analysis and western blot analysis of TRIM8 among two normal cell lines and six patient‐derived GBM neurosphere cells. NHNP = normal human neural progenitor cell. (I) Immunocytochemical (ICC) staining of TRIM8 in neurosphere cells showing nuclear expression of TRIM8. TRIM8 was labeled in GFP, and nuclei were counterstained with DAPI (blue). Scale bar = 10 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28100038), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Rt
• Applications: ICC/IF, IHC, Mycoplasma
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

TRIM8 Antibody - BSA Free Summary

• Immunogen: This antibody was developed against Recombinant Protein corresponding to amino acids: QDIEDQLYKLESDKRLVEEKVNQLKEEVRLQYEKLHQLLDEDLRQTVEVLDKAQAKFCSENAAQALHLGERMQEAKKLLGS
• Predicted Species: Mouse (99%), Rat (99%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: TRIM8
• Purity: Affinity purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 0.25-2 ug/ml
  • Immunohistochemistry 1:50 - 1:200
  • Immunohistochemistry-Paraffin 1:50 - 1:200
  • Knockdown Validated
• Application Notes: IHC-Paraffin, HIER pH 6 retrieval is recommended. ICC/IF, Fixation Permeabilization: Use PFA/Triton X-100.
• Control Peptide:

  TRIM8 Recombinant Protein Antigen (NBP1-89776PEP)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH 7.2) and 40% Glycerol
• Preservative: 0.02% Sodium Azide
• Purity: Affinity purified

Alternate Names for TRIM8 Antibody - BSA Free

• EC 6.3.2.-
• GERP
• GERPRING finger protein 27glioblastoma expressed ring finger protein
• Glioblastoma-expressed RING finger protein
• RNF27probable E3 ubiquitin-protein ligase TRIM8
• tripartite motif containing 8
• tripartite motif protein TRIM8
• tripartite motif-containing 8
• Tripartite motif-containing protein 8

Background

TRIM8 is a member of the tripartite motif (TRIM) family. The TRIM motif includes three zinc-binding domains, a RING, a B-box type 1 and a B-box type 2, and a coiled-coil region. This protein localizes to nuclear bodies. Its structure is similar to some tumor suppressor proteins and its gene maps to a locus thought to contain tumor suppressor genes.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for TRIM8 Antibody (NBP1-89776)(1)

Publication using NBP1-89776

• Zhang C, Mukherjee S, Tucker-Burden C et al. TRIM8 regulates stemness in glioblastoma through PIAS3-STAT3 Mol Oncol 2017-02-15 [PMID: 28100038] (WB, FLOW, Human)

Bioinformatics

• Gene Symbol: TRIM8
• Entrez:
  • 81603(Human)
• Uniprot:
  • Q9BZR9(Human)