Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-19058 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Ferritin Heavy Chain Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with canine based on sequence homology. This antibody is tested in Peptide ELISA: antibody detection limit dilution 4,000.
- Reactivity
- Human
- Host
- Goat
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot staining of Human Placenta lysate using Product # PA5-19058 at a concentration of 0.05 µg/mL, the primary antibody incubation was 1 hour and the detection method was chemiluminescence.
- 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 # PA5-19058, 1ug/ml) and Rabbit Anti-Goat IgG Secondary Antibody, HRP conjugate (Product # A27014, 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Ferritin Heavy Chain Goat Polyclonal Antibody (Product # PA5-19058) and a 20kDa band corresponding to Ferritin Heavy Chain was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1) and PC-3 (Lane 2) 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 (1ug/ml) and detected by chemiluminescence with Rabbit Anti-Goat IgG Secondary Antibody, HRP conjugate (Product # A27014, 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 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 # PA5-19058) 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
- Immunohistochemical staining of paraffin embedded of Human Kidney using Product # PA5-19058 at a concentration of 4 µg/mL. The tissue was processed by steamed antigen retrieval with citrate buffer pH 6 and stained with AP.