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LearnMA5-15141
antibody from Invitrogen Antibodies
Targeting: RPS6KB1
p70(S6K)-alpha, PS6K, S6K, S6K1, STK14A
Western blotAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
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- Product number
- MA5-15141 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- p70 S6 Kinase Monoclonal Antibody (T.527.6)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody.
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- T.527.6
- Vial size
- 100 μL
- Concentration
- 48 μg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-p70 S6 Kinase Monoclonal Antibody (T.527.6) (Product # MA5-15141) and a 57, 59 kDa band corresponding to RPS6KB1 was observed across the panel tested. Whole cell extracts (30 µg lysate) of A549 (Lane 1), LNCaP (Lane 2), A-431 (Lane 3), MCF7 (Lane 4), HEK-293 (Lane 5), PC-3 (Lane 6), Mouse Brain (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX), 10 well. 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 Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of RPS6KB1 was performed by loading 30 µg of HeLa WT Control (Lane 1), HeLa Cas9 Control (Lane 2) and HeLa- RPS6KB1 knockout (Lane 3) whole cell extracts. The blot was probed with Anti-p70 S6 Kinase Monoclonal Antibody (T.527.6) Antibody (Product # MA5-15141, 1:1000 dilution) and Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to RPS6KB1.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of RPS6KB1 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 45 minutes at room temperature. The cells were labeled with p70 S6 Kinase Monoclonal Antibody (T.527.6) (Product # MA5-15141) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790, 1:3000 dilution), 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 cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
