- MA1-12607 - Invitrogen Antibodies FYN antibody

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Journal of immunology research 2015;2015:395371

Boggs JM, Homchaudhuri L, Ranagaraj G, Liu Y, Smith GS, Harauz G
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DeBruin LS, Haines JD, Wellhauser LA, Radeva G, Schonmann V, Bienzle D, Harauz G
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Chen M, Wang Y, Liu Y, Hou XY, Zhang QG, Meng FJ, Zhang GY
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Chen M, Hou X, Zhang G
Neuroscience letters 2003 Mar 13;339(1):29-32

Chen M, Hou X, Zhang G
Neuroscience letters 2003 Mar 13;339(1):29-32

Hou XY, Zhang GY, Zong YY
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Hou XY, Zhang GY, Yan JZ, Liu Y
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Hou XY, Zhang GY, Yan JZ, Chen M, Liu Y
Brain research 2002 Nov 15;955(1-2):123-32

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of HEK 293 (Lane 1), MCF7 (Lane 2) and Mouse Liver (Lane 3). The blot was probed with Anti-Fyn Mouse Monoclonal Antibody (Product # MA1-12607, 1-3 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A 60 kDa band corresponding to Fyn was observed across the cell lines and tissue tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of HEK 293 (Lane 1), MCF7 (Lane 2) and Mouse Liver (Lane 3). The blot was probed with Anti-Fyn Mouse Monoclonal Antibody (Product # MA1-12607, 1-3 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A 60 kDa band corresponding to Fyn was observed across the cell lines and tissue tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Fyn in formalin-fixed, paraffin-embedded human lymphoma using a Fyn monoclonal antibody (Product # AHO0482). Detection was performed with a peroxidase-conjugate and AEC chromogen. Note cytoplasmic staining of tumor cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 Fyn detection in stimulated human PBMC. (a) PBMC were treated with PHA-P and hTip-CSKH during 2 h, lysed, and the protein extract resolved by SDS-PAGE. Western blot was performed with an anti-Fyn specific antibody. PHA-P was used as positive control, which induces the appearance of a second band of 56 kDa. (b) Quantification of the Fyn p59 and p56 bands. The intensity of each Fyn band was normalized to the loading control beta -actin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation and Western blot of Fyn was performed using mouse spleen cell lysates. Cells were lysed using ice cold IP lysis/wash buffer and pre-cleared using 50 mL of bead slurry per mL of cell lysate. Antigen-antibody complexes were formed by incubating 0.5 mL pre-cleared cell lysate on ice for 3hrs with 8-15 µg of Fyn monoclonal antibody (Product # AHO0482) crosslinked to Protein A/G plus agarose. The immune complexes were eluted using 60 µL sample buffer boiled at 95°C for 5 min and loaded onto an SDS-PAGE gel; Input (lane 1) and Jurkat IP (Lane 2). The membrane was probed with a Fyn monoclonal antibody (Product # AHO0482) at a dilution of 3 µg/mg of lysate followed by detection using an HRP-conjugated goat anti-mouse IgG + IgM (H+L) cross-adsorbed secondary antibody. Chemiluminescent detection was performed using an exposure time of 10m, resulting in a ~60 kDa band on input and IP lysates.

MA1-12607