Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Immunocytochemistry [1]
- Other assay [3]
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- Product number
- MA5-14735 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Ferritin Monoclonal Antibody (523)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- MA5-14735 detects Ferritin in ELISA, IP, IF, and RIA applications and shows reactivity to human samples. The MA5-14735 Immunogen is human liver ferritin. MA5-14735 detects ferritin which has a predicted molecular weight of approximately 19kDa. Product MA514735 is a smaller package size of MIF2503 (formerly sold as a Seradyn product).
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 523
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Ferritin (FTL, green) in HepG2 cells and MCF7 cells. The cells were fixed with formalin for 15 minutes, permeabilized with 0.1% Triton X-100 in TBS for 10 minutes, and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were stained with a Ferritin monoclonal antibody (Product # MA5-14735), at a dilution of 1:200 for at least 1 hour at room temperature, and then incubated with a DyLight 488 goat anti-mouse secondary antibody (Product # 35502) at a dilution of 1:500 for 30 minutes at room temperature (both panels, green). F-Actin (both panels, red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (both panels, blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight at 20X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Direct ELISA analysis of Ferritin was performed by coating wells of a 96-well plate with 100 µL per well of Human Ferritin or BSA diluted in carbonate/bicarbonate buffer (Product # 28382), starting at a concentration of 1 µg/mL and serially diluting 2-fold to a concentration of 2 ng/mL, overnight at 4C. Wells of the plate were washed, blocked with StartingBlock blocking buffer (Product # 37538), and incubated with 100 µL per well of a mouse anti-ferritin monoclonal antibody (Product # MA5-14735) at a concentration of 1 µg/mL for 1 hour at room temperature. The plate was washed, then incubated with 100 µL per well of an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) at a dilution of 1:25000 for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 10 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550 nm.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Sandwich ELISA of Ferritin was performed by coating wells of a 96-well plate with 100 µL of an Ferritin mouse monoclonal antibody (Product # MA5-14735) diluted to a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 100 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 28382) for 2 hours, and 100 µL of recombinant human ferritin light polypeptide (FTL) standards ranging from 1.6-1000 ng/mL (A) or 100 µL of diluted normal human serum or diluted normal human plasma (B) were incubated for 1 hour at room temperature. The plate was washed with 1X TBST (Product # 28360), and 100 µL per well of an biotin-conjugated ferritin rabbit polyclonal antibody (Product # PA1-26882) diluted to a concentration of 1 µg/mL was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of a Streptavidin-HRP (Product # 21126) was incubated for 30 minutes. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Sandwich ELISA of Ferritin was performed by coating wells of a 96-well plate with 100 µL of an Ferritin duck polyclonal antibody (Product # D0107-2D) diluted to a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 100 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 28382) for 2 hours, and 100 µL of recombinant human ferritin light polypeptide (FTL) standards ranging from 1.6-1000 ng/mL were incubated for 1 hour at room temperature. The plate was washed with 1X TBST (Product # 28360), and 100 µL per well of an ferritin mouse monoclonal antibody (Product # MA5-14735) diluted to a concentration of 1 µg/mL was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of a Streptavidin-HRP (Product # 21126) was incubated for 30 minutes. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.