Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-57858 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- RIF1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Immunogen sequence: FLPKAKQREG TFSKSDSEKI VNGTKRSSRR AGKAEQTGNK RSKPLMRSEP EKNTEESVEG IVVLENNPPG LLNQTECVSD NQVHLSESTM EHD Highest antigen sequence identity to the following orthologs: Mouse - 49%, Rat - 52%.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.3 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Telomere-associated protein RIF1 was achieved by transfecting K-562 with Telomere-associated protein RIF1 specific siRNAs (Silencer® select Product # S30376, S30378). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the Telomere-associated protein RIF1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with RIF1 Polyclonal Antibody (Product # PA5-57858, 1:500 ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). 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 Telomere-associated protein RIF1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-RIF1 Polyclonal Antibody (Product # PA5-57858) and a 274 kDa band corresponding to Telomere-associated protein RIF1 was observed. Nuclear enriched extracts (30 µg lysate) of K-562 (Lane 1) and HeLa (Lane 2) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). 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 (1:500) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). RIF1, being a telomere binding protein, interacts with a wide range of proteins which are involved in maintaining telomere stability and in controlling the replication process. Such interactions have high affinity and they are difficult to be dissociated under denaturing conditions. Hence, a RIF1 antibody detects multiple bands in Western Blot.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical staining of RIF1 in human testis tissue shows strong nuclear positivity. Samples were probed using a RIF1 Polyclonal Antibody (Product # PA5-57858).