Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Other assay [1]
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Validation data
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- Product number
- PA5-17453 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- c-Cbl Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 14.5 µg/mL
- Storage
- -20°C
Submitted references c-Cbl Acts as an E3 Ligase Against DDA3 for Spindle Dynamics and Centriole Duplication during Mitosis.
Gwon D, Hong J, Jang CY
Molecules and cells 2019 Dec 31;42(12):840-849
Molecules and cells 2019 Dec 31;42(12):840-849
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of c-Cbl in lysates from RAW and COS cells using c-Cbl polyclonal antibody (Product # PA5-17453).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of c-Cbl was achieved by transfecting SH-SY5Y cells with c-Cbl specific siRNA (Silencer® select Product # s2477). Western blot analysis (Fig. a) was performed using whole cell extracts from the c-Cbl knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with c-Cbl Polyclonal Antibody (Product # PA5-17453, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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 c-Cbl.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-c-Cbl polyclonal Antibody (Product # PA5-17453) and a 110 kDa band corresponding to c-Cbl was observed in Mouse testis, Rat testis, THP-1, BeWo, Daudi and SH-SY5Y but was absent in Mouse Heart and Rat heart, as reported. Tissue extracts (30 µg lysate) of Mouse testis (Lane 1), Rat testis (Lane 2), Mouse heart (Lane 3), Rat heart (Lane 4), and whole cell extracts (30 µg lysate) of THP-1 (Lane 5), BeWo (Lane 6), Daudi (Lane 7) and SH-SY5Y (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 (1:1000 dilution) 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
- Immunofluorescent analysis of c-Cbl in paraformaldehyde-fixed RAW cells, using a c-Cbl polyclonal antibody (Product # PA5-17453).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of c-Cbl in paraformaldehyde-fixed RAW cells, using a c-Cbl polyclonal antibody (Product # PA5-17453).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Fig. 1 c-Cbl interacts with DDA3 and acts as a mitotic regulator (A) The DDA3 complex was purified from mitotic cells and analyzed by mass spectrometry. A peptide of c-Cbl was identified three times. (B) Twenty-eight hours after transfection of the EV or HA-c-Cbl plasmid, HeLa cells were harvested and subjected to immunoprecipitation and western blotting with the indicated antibodies. p38MAPK served as a loading control. EV, empty vector. (C and D) HeLa cells were synchronized by a double thymidine block (C) or thymidine-nocodazole block (D), placed into fresh media, and harvested at the indicated times. Cell lysates were analyzed by western blotting with the indicated antibodies. AS, unsynchronized cells. (E) HeLa cells were transfected with control (siControl) or c-Cbl-specific siRNAs (siCbl-A and siCbl-B). Seventy-two hours after siRNA transfection, the transfected cells were harvested and lysed to measure protein levels by western blotting with the indicated antibodies. (F and G) Seventy-two hours after siRNA transfection, HeLa cells were fixed with MeOH and stained with antibodies as indicated. Images are maximum projections from Z-stacks of representative cells stained for c-Cbl (green), beta-tubulin (red), and DNA (blue). The number of metaphase cells with unaligned chromosomes was quantified and plotted (G) (n = 300 metaphase cells from three independent experiments). (H) Seventy-two hours after siRNA transfection, HeLa cells expressing GFP-Histone H2B cells were imag