Ferritin Recombinant Rabbit Monoclonal Antibody [SC0620]
cat.: ET1610-78
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Zebrafish, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P
Clonality: Monoclonal
Clone number: SC0620
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 21 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human FTH1 aa 58-99 / 183.
Positive control: 293T cell lysate, RAW264.7 cell lysate, mouse liver tissue lysate, mouse brain tissue lysate, rat liver tissue lysate, rat brain tissue lysate, zebrafish tissue lysates, hybrid fish (crucian-carp) liver tissue lysates, SGC-7901, MCF-7, SH-SY5Y, human spleen tissue.
Subcellular location: Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
1:2,000-1:5,000
1:50-1:100
1:50-1:100
1:50-1:200
Uniprot #: SwissProt: P02794 Human | P09528 Mouse | P19132 Rat
Alternative names: Cell proliferation-inducing gene 15 protein Ferritin H subunit Ferritin heavy chain Ferritin heavy polypeptide 1 Ferritin L subunit Ferritin, heavy polypeptide FRIH_HUMAN FTH FTH1 FTL
Images
ET1610-78_1.jpg Fig1: Western blot analysis of Ferritin on different lysates with Rabbit anti-Ferritin antibody (ET1610-78) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

Lane 1: 293T cell lysate (30 µg/Lane)
Lane 2: RAW264.7 cell lysate (15 µg/Lane)
Lane 3: Mouse liver tissue lysate (15 µg/Lane)
Lane 4: Mouse brain tissue lysate (15 µg/Lane)
Lane 5: Rat liver tissue lysate (15 µg/Lane)
Lane 6: Rat brain tissue lysate (30 µg/Lane)

Predicted band size: 21 kDa
Observed band size: 21 kDa

Exposure time: 3 minutes 20 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-78) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1610-78_2.jpg Fig2: All lanes: Western blot analysis of Ferritin with anti-Ferritin antibody (ET1610-78) at 1:1,000 dilution.
Lane 1/2: Wild-type SHSY5Y whole cell lysate (10 µg).
Lane 3/4: Ferritin fragment 1 knockout SHSY5Y whole cell lysate (10 µg).
Lane 5/6: Ferritin fragment 1 knockout SHSY5Y whole cell lysate (10 µg).

ET1610-78 was shown to specifically react with Ferritin in wild-type SHSY5Y cells. No band was observed when Ferritin knockout sample was tested. Wild-type and Ferritin knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1610-78, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
ET1610-78_3.jpg Fig3: Immunocytochemistry analysis of SGC-7901 cells labeling Ferritin with Rabbit anti-Ferritin antibody (ET1610-78) at 1/100 dilution.

Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Ferritin antibody (ET1610-78) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1610-78_4.jpg Fig4: ICC staining of Ferritin in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-78, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1610-78_5.jpg Fig5: ICC staining of Ferritin in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-78, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1610-78_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Ferritin antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-78, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.