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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [5]
- Immunocytochemistry [1]
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- Product number
- MA1-23248 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- JAB1 Monoclonal Antibody (8H8.5)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Recommended positive controls: 293 cell lysate.
- Reactivity
- Human, Mouse, Rat, Drosophila
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 8H8.5
- Vial size
- 100 µL
- Concentration
- 1 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
- Western blot analysis of JAB1 in HeLa whole cell lysate using a JAB1 monoclonal antibody (Product # MA1-23248).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using JAB1 Monoclonal Antibody (8H8.5) (Product # MA1-23248). Part of a customer review using JAB1 Monoclonal Antibody (8H8.5) (Product # MA1-23248).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using JAB1 Monoclonal Antibody (8H8.5) (Product # MA1-23248). Customer Review using JAB1 Monoclonal Antibody (8H8.5) (Product # MA1-23248) - D10 T cells were lysed in IP buffer (20mM Tris-HCl, pH 7.5, 50mM NaCl, 50mM NaF, 10mM -glycerophosphate, 1% Triton X-100, 1mM PMSF).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of JAB1 was achieved by transfecting SH-SY5Y cells with JAB1 specific siRNAs (Silencer® select Product # s21627, s21628). Western blot analysis (Fig. a) was performed using whole cell extracts from the SH-SY5Y knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with JAB1 Monoclonal Antibody (8H8.5) (Product # MA1-23248, 0.5 µg/ml) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP (Product # A28177, 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 JAB1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-JAB1 Monoclonal Antibody (Product # MA1-23248) and a ~37 kDa band corresponding to JAB1 was observed across the cell lines and tissues tested along with a 25 kDa band (*) corresponding to light chain of IgG in mouse tissues. Whole cell extracts (30 ug lysate) of A-431 (Lane 1), A549 (Lane 2), HEK-293 (Lane 3), HeLa (Lane 4), MCF-7 (Lane 5), HT-29 (Lane 6), SW-480 (Lane 7), HCT 116 (Lane 8), Mouse Lung (Lane 9), Rat Lung (Lane 10) and Mouse Spleen (Lane 11) 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 (1:2000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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 JAB1 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 JAB1 Monoclonal Antibody (Product # MA1-23248) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). 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.
