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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunohistochemistry [2]
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- Product number
- PA5-19127 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- FMRP Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with rat based on sequence homology. This antibody is tested in Peptide ELISA: antibody detection limit dilution 32,000.
- 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 HeLa cell lysate using Product # PA5-19127 at a concentration of 0.3 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 FMRP was achieved by transfecting HEK-293 cells with FMRP specific siRNAs (Silencer® select Product # s5316, s5317). Western blot analysis (Fig. a) was performed whole cell extracts from the HEK-293 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with FMRP Polyclonal Antibody (Product # PA5-19127, 0.25 µg/mL) and Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (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 FMRP (71 kDa isoform).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-FMRP Goat Polyclonal Antibody (Product # PA5-19127) and bands varying between 29-71 kDa band corresponding to FMRP were observed across cell lines tested. Whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), U-87 MG (Lane 2), HeLa (Lane 3), A549 (Lane 4), SH-SY5Y (Lane 5) and Hep G2 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 (0.5 µg/mL) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (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
- Immunohistochemical analysis of FMRP in paraffin embedded human cortex using a FMRP polyclonal antibody (Product #PA5-19127) at a concentration of 2.5 µg/mL. Steamed antigen retrieval was performed with pH 6 buffered citrate. Samples were then stained with alkaline phosphatase.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of FMRP in paraffin embedded human cortex using a FMRP polyclonal antibody (Product #PA5-19127) at a concentration of 2.5 µg/mL. Steamed antigen retrieval was performed with pH 6 buffered citrate. Samples were then stained with alkaline phosphatase.
