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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Flow cytometry [2]
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- Product number
- 701396 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SRC Recombinant Rabbit Monoclonal Antibody (14H2L20)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 14H2L20
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
Submitted references Dual clathrin and integrin signaling systems regulate growth factor receptor activation.
Alfonzo-Méndez MA, Sochacki KA, Strub MP, Taraska JW
Nature communications 2022 Feb 16;13(1):905
Nature communications 2022 Feb 16;13(1):905
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SRC was performed by loading 30 µg of MDA-MB-231 (lane1), MCF 7 (lane2), Jurkat (lane3), A431 (lane4), U2OS (lane5), Hep G2 (lane 6), HEK-293 (lane 7) and HeLa (lane 8) cell lysate using Novex®NuPAGE®4-12 % Bis-Tris gel (Product # NP0322PK2), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800), and iBlot® Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature. SRC was detected at ~ 59 kDa using SRC Recombinant Rabbit Monoclonal Antibody (Product # 701396) at 1-3 µg/mL in 5% skim milk at 4°C overnight on a rocking platform. Goat anti-Rabbit IgG - HRP Secondary Antibody (Product # G-21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SRC in whole cell extracts from A431, HepG2, and U20S (lanes 1-3 respectively) using a SRC recombinant rabbit monoclonal antibody (Product # 701396) at a dilution of 1 µg/mL. Detection was performed using an HRP-conjugated goat anti-rabbit secondary antibody followed by chemiluminescence (ECL). Results show a band at ~59kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SRC was performed by loading 30 ug of HeLa control (Lane 1), HeLa- SRC knockout (Lane 2) edigene lysates. The blot was probed with SRC Antibody (14H2L20), Recombinant Rabbit Monoclonal (Product # 701396, 1 µg/ml) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to SRC.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of SRC in HeLa cells using a SRC recombinant rabbit monoclonal antibody (Product # 701396) followed by detection using an Alexa Fluor 488-conjugated goat anti-rabbit secondary antibody (green) (Image A). Nuclei were stained using DAPI (Image B) and actin stained with Alexa Fluor 594 phalloidin (red) (image C). Image D is a composite image showing cytoplasmic and nuclear localization of SRC.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of SRC was done on A431 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Tritonª X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with ABfinityª SRC Recombinant Rabbit Monoclonal Antibody (701396, red histogram) or with rabbit isotype control (pink histogram) at 3-5 µg/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor¨ 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune¨ Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of SRC in HeLa cells using a SRC recombinant rabbit monoclonal antibody (Product # 701396). Cells were fixed and permeabilized using FIX & PERM (Product # GAS-004) reagent, and detection was performed using an Alexa Fluor 488 goat anti-rabbit IgG (right peak) compared to an isotype control (middle peak, black) and a control without primary antibody (left peak, blue).
