- PA5-27500 - Invitrogen Antibodies FTH1 antibody

Submitted references Mutations in LRRK2 linked to Parkinson disease sequester Rab8a to damaged lysosomes and regulate transferrin-mediated iron uptake in microglia.

Mamais A, Kluss JH, Bonet-Ponce L, Landeck N, Langston RG, Smith N, Beilina A, Kaganovich A, Ghosh MC, Pellegrini L, Kumaran R, Papazoglou I, Heaton GR, Bandopadhyay R, Maio N, Kim C, LaVoie MJ, Gershlick DC, Cookson MR
PLoS biology 2021 Dec;19(12):e3001480

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Ferritin Heavy Chain using 30 µg of A549 lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a Ferritin Heavy Chain polyclonal antibody (Product # PA5-27500) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of Ferritin Heavy Chain was performed by separating 30 µg of human tissue extract by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a Ferritin Heavy Chain Polyclonal Antibody (Product # PA5-27500) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of Ferritin Heavy Chain was performed by separating 50 µg of mouse tissue extract by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a Ferritin Heavy Chain Polyclonal Antibody (Product # PA5-27500) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of Ferritin Heavy Chain was performed by separating 50 µg of rat tissue extract by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a Ferritin Heavy Chain Polyclonal Antibody (Product # PA5-27500) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of Ferritin Heavy Chain was performed by separating 30 µg of whole cell extract by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a Ferritin Heavy Chain Polyclonal Antibody (Product # PA5-27500) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of Ferritin Heavy Chain was performed by separating 30 µg of whole cell extract by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a Ferritin Heavy Chain Polyclonal Antibody (Product # PA5-27500) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Ferritin Heavy Chain Polyclonal Antibody (Product # PA5-27500). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with Ferritin Heavy Chain Polyclonal Antibody (Product # PA5-27500) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Ferritin Heavy Chain Polyclonal Antibody (Product # PA5-27500). Various tissue extracts (50 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with Ferritin Heavy Chain Polyclonal Antibody (Product # PA5-27500) diluted at 1:20,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Ferritin Heavy Chain was achieved by transfecting HeLa cells with Ferritin Heavy Chain specific siRNAs (Silencer® select Product # s21066, s21067). Western blot analysis (Fig. a) was performed whole cell extracts from the HeLa knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Ferritin Heavy Chain Polyclonal Antibody (Product # PA5-27500, 1:2000 dilution) and Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 0.25 ug/ml, 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 Ferritin Heavy Chain.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Ferritin Heavy Chain Rabbit Polyclonal Antibody (Product # PA5-27500) and a 20kDa band corresponding to Ferritin Heavy Chain was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), PC-3 (Lane 2), Mouse Ovary (Lane 3), Mouse Spleen (Lane 4), Rat Ovary (Lane 5) and Rat Spleen (Lane 6) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). 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 (1:2000 dilution) and detected by chemiluminescence with Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 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: Immunocytochemistry-Immunofluorescence analysis of Ferritin Heavy Chain was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: Ferritin Heavy Chain Polyclonal Antibody (Product # PA5 27500) diluted at 1:500. Red: Phalloidin, a cytoskeleton marker. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Ferritin Heavy Chain 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 Ferritin Heavy Chain Polyclonal Antibody (Product # PA5-27500) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear and cytoplasmic 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: Immunohistochemistry (Paraffin) analysis of Ferritin Heavy Chain was performed in paraffin-embedded mouse liver tissue using Ferritin Heavy Chain Polyclonal Antibody (Product # PA5-27500) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry (Paraffin) analysis of Ferritin Heavy Chain was performed in paraffin-embedded rat liver tissue using Ferritin Heavy Chain Polyclonal Antibody (Product # PA5-27500) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 10.1371/journal.pbio.3001480.g008 Fig 8 Inflammation induces ferritin accumulation in microglia in G2019S LRRK2 knock-in mice. FTH and Tf were stained and visualized in collected brains, 72 hours after post-intrastriatal injections of LPS. G2019S LRRK2 mice exhibited higher levels of FTH across the striatum compared to WT and Lrrk2 KO mice while Tf was not altered significantly in the knock-in (A, B, C) ( N = 4 WT mice, 4 G2019S and 2 Lrrk2 KO; B: two-way ANOVA, Tukey post hoc, Genotype: ** P = 0.0017, F (2, 7) = 18.27, Distance from injection site: **** P < 0.0001, F (8, 56) = 50.25; C: two-way ANOVA, Tukey post hoc, Genotype: P = 0.1040, F (2, 7) = 3.182, Distance from injection site: **** P < 0.0001, F (8, 56) = 29.17). Microglia are positive for FTH in both WT and G2019S LRRK2 cohorts (D). Similar percentages of FTH-positive microglia were observed between the WT and G2019S LRRK2 groups (E). [SEM bars are shown]. The underlying data can be found in S1 Data . FTH, ferritin heavy chain; KO, knockout; LPS, lipopolysaccharide; LRRK2, leucine-rich repeat kinase 2; Tf, transferrin; WT, wild-type.

PA5-27500

Antibody data