ET1610-78

Ferritin Heavy Chain 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, SGC-7901, human spleen tissue, rat spleen tissue.
Subcellular location:	Cytoplasm.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P	1:2,000-1:5,000 1:50-1:100 1:200 1:1,000
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

Fig1: Western blot analysis of Ferritin Heavy Chain on different lysates with Rabbit anti-Ferritin Heavy Chain 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.

Fig2: All lanes: Western blot analysis of Ferritin Heavy Chain with anti-Ferritin Heavy Chain antibody (ET1610-78) at 1:1,000 dilution.
Lane 1/2: Wild-type SHSY5Y whole cell lysate (10 µg).
Lane 3/4: Ferritin Heavy Chain fragment 1 knockout SHSY5Y whole cell lysate (10 µg).
Lane 5/6: Ferritin Heavy Chain fragment 1 knockout SHSY5Y whole cell lysate (10 µg).

ET1610-78 was shown to specifically react with Ferritin Heavy Chain in wild-type SHSY5Y cells. No band was observed when Ferritin Heavy Chain knockout sample was tested. Wild-type and Ferritin Heavy Chain 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.

	Fig3: Immunocytochemistry analysis of SGC-7901 cells labeling Ferritin Heavy Chain with Rabbit anti-Ferritin Heavy Chain 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 Heavy Chain 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Ferritin Heavy Chain antibody (ET1610-78) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1610-78) at 1/1,000 dilution for 1 hour 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Ferritin Heavy Chain antibody (ET1610-78) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1610-78) at 1/1,000 dilution for 1 hour 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.

Fig6: Western blot analysis of Ferritin Heavy Chain on different lysates with Rabbit anti-Ferritin Heavy Chain antibody (ET1610-78) at 1/1,000 dilution.

Lane 1: FTL recombinant protein, 20ng/Lane
Lane 2: FTH1 recombinant protein, 20ng/Lane

Exposure time: 20 seconds; ECL: K1801;

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 1/1,000 dilution was 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.