480009

antibody from Invitrogen Antibodies

Targeting: FBL FIB, FLRN, Nop1, RNU3IP1

Immunohistochemistry

Antibody data

Submit

Validation data

Product number: 480009 - Provider product page
Provider: Invitrogen Antibodies
Product name: Fibrillarin Monoclonal Antibody (38F3)
Antibody type: Monoclonal
Antigen: Other
Reactivity: Human, Mouse, Rat, Bovine, Chicken/Avian, Drosophila, Porcine, Yeast
Host: Mouse
Isotype: IgG
Antibody clone number: 38F3
Vial size: 100 µL
Storage: -20° C, Avoid Freeze/Thaw Cycles

FBL protein structure - 480009 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of HeLa lysate showing specific immunolabeling of the ~ 34k fibrillarin protein.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Fibrillarin was achieved by transfecting HeLa with Fibrillarin specific siRNAs (Silencer® select Product # s4820, s4822). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the Fibrillarin knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Fibrillarin Monoclonal Antibody (38F3) (Product # 480009, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Fibrillarin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of Fibrillarin in HeLa cell lysate showing specific immunolabeling of a ~34 kDa band corresponding to Fibrillarin monoclonal antibody (Product # 480009).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Fibrillarin Monoclonal Antibody (38F3) (Product # 480009) and a 38 kDa band corresponding to Fibrillarin was observed across cell lines tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), PANC-1 (Lane 3), U-2 OS (Lane 4), A-431 (Lane 5), MCF-7 (Lane 6), HT-29 (Lane 7), HEK-293 (Lane 8) and PC-12 (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Fibrillarin was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Fibrillarin Mouse Monoclonal Antibody (38F3) (Product # 480009) at 1:250 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nucleolar localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of Fibrillarin in HeLa cells. Showing strong prominent labeling of the nucleoli in the nucleus with Fibrillarin monoclonal antibody (Product # 480009) using a dilution of 1:100 (green). Colabeled with Anti-Vimentin antibody using a dilution of 1:1,000 (red) showing the cytoplasmic intermediated filaments, and nuclear staining was done with DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of Fibrillarin in human neuroblastoma SH-SY5Y cells. Showing strong prominent labeling of the nucleoli in the nucleus with Fibrillarin monoclonal antibody (Product # 480009) with a dilution of 1:500 (green). A nuclear staining was done with DAPI (blue) which makes the fibrillarin marker appear pale blue.