FBL Monoclonal antibody

FBL Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Conjugate

Unconjugated

Publications

5

CloneNo.

3A9E8

Cat no : 66985-1-Ig

Synonyms

FLRN, fibrillarin, FIB1, FIB, EC:2.1.1.-

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Validation Data Gallery

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  WB analysis using 66985-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66985-1-Ig (FBL antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Alpha Tubulin Monoclonal antibody (HRP-66031) as loading control.

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  WB analysis using 66985-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66985-1-Ig (FBL antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 66985-1-Ig

  IP result of anti-FBL (IP:66985-1-Ig, 5ug; Detection:66985-1-Ig 1:1000) with HeLa cells lysate 1600 ug.

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  IHC staining of human liver cancer using 66985-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66985-1-Ig (FBL antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human liver cancer using 66985-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66985-1-Ig (FBL antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 66985-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using FBL antibody (66985-1-Ig, Clone: 3A9E8 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L), Alpha Tubulin antibody (11224-1-AP, red).

• 

  IF Staining of HeLa using 66985-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using FBL antibody (66985-1-Ig, Clone: 3A9E8 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L), Alpha Tubulin antibody (11224-1-AP, red).

• 

  IF Staining of HepG2 using 66985-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using FBL antibody (66985-1-Ig, Clone: 3A9E8 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

• 

  FC experiment of HepG2 using 66985-1-Ig

  1X10^6 HepG2 cells were intracellularly stained with 0.4 ug Anti-Human FBL (66985-1-Ig, Clone:3A9E8) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG1 Isotype Control (MOPC-21) (65124-1-Ig, Clone: MOPC-21) (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, HepG2 cells, Jurkat cells, HSC-T6 cells, NIH/3T3 cells, 4T1 cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human liver cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells, HepG2 cells
• Positive FC (Intra) detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 4 publications below
• IF: See 3 publications below

Product Information

66985-1-Ig targets FBL in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: FBL fusion protein Ag8841
• Full Name: fibrillarin
• Calculated Molecular Weight: 321 aa, 34 kDa
• Observed Molecular Weight: 34 kDa
• GenBank Accession Number: BC019260
• Gene Symbol: FBL
• Gene ID (NCBI): 2091
• RRID: AB_2882303
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Fibrillarin (Fbl) is an essential nucleolar protein that plays an essential role in ribosome biogenesis and more particularly in the methylation of ribosomal RNAs and rDNA histones. FBL is highly expressed in various cancers and correlates with poor survival outcomes in cancer patients,such as hepatocellular carcinoma(PMID: 33376525). Knockdown of FBL sensitizes tumor cells and xenografts to DNA crosslinking agents, and leads to homologous recombination-mediated DNA repair defects(PMID: 37489617).

Publications

Species	Application	Title
human	WB,IF	Gastroenterology; Nucleolar HEATR1 upregulated by mTORC1 signaling promotes hepatocellular carcinoma growth by dominating ribosome biogenesis and proteome homeostasis; Authors - Xiao-Mei Yang
human	WB	BMC Biol; A pair of long intergenic non-coding RNA LINC00887 variants act antagonistically to control Carbonic Anhydrase IX transcription upon hypoxia in tongue squamous carcinoma progression; Authors - Tao Shen
human	WB	Thorac Cancer; LncRNA FAM83A-AS1 promotes lung adenocarcinoma progression by enhancing the pre-mRNA stability of FAM83A.; Authors - Wenyi Wang
human	IF	Cell Death Discov; Protein expression of nucleolar protein 12 in the retina and its implication in protection of retina from UV irradiation damage; Authors - Jingtao Liu
human	WB,IF	Nat Commun; NAP-seq reveals multiple classes of structured noncoding RNAs with regulatory functions; Authors - Shurong Liu