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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [6]
- Immunohistochemistry [2]
- Other assay [1]
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- Product number
- PA5-21494 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- AMPK alpha-2 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, HeLa, HepG2, NIH-3T3.
- Concentration
- 0.96 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of AMPK alpha 2 using A) 30 µg A431 whole cell lysate (B) 30 µg HeLa whole cell lysate and C) 30 µg A375 whole cell lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with an AMPK alpha 2 polyclonal antibody (Product # PA5-21494) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of AMPK-alpha-2 was performed by separating 30 µg of various whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a AMPK alpha-2 Polyclonal Antibody (Product # PA5-21494) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of AMPK alpha-2 was achieved by transfecting MDA-MB-231 cells with AMPK alpha-2 specific validated siRNAs (Silencer® select Product # s11057, s11056). Western blot analysis (Fig. a) was performed using whole cell extracts from the AMPK alpha-2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with AMPK alpha-2 Polyclonal Antibody (Product # PA5-21494, 1:500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/mL, 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 PA5-21494.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of MDA-MB-231 (Lane 1), HEK-293 (Lane 2), Hep G2 (Lane 3) and tissue extracts (30 µg lysate) of Mouse skeletal muscle (Lane 4) and Rat heart (Lane 5). The blot was probed with Anti-AMPK alpha-2 Polyclonal Antibody (Product # PA5-21494, 1:500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 63 kDa band corresponding to AMPK alpha-2 was observed across the cell lines and tissue extracts tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of AMPK-alpha-2 was performed by separating 30 µg of NIH-3T3 lysates by 7.5% SDS PAGE. Proteins were transferred to a membrane and probed with a AMPK alpha-2 Polyclonal Antibody (Product # PA5-21494) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of AMPK-alpha-2 was performed by separating 30 µg of various whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a AMPK alpha-2 Polyclonal Antibody (Product # PA5-21494) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry (Paraffin) analysis of AMPK alpha-2 was performed in paraffin-embedded mouse muscle tissue using AMPK alpha-2 Polyclonal Antibody (Product # PA5-21494) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry (Paraffin) analysis of AMPK alpha-2 was performed in paraffin-embedded rat muscle tissue using AMPK alpha-2 Polyclonal Antibody (Product # PA5-21494) at a dilution of 1:500.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- AMPK alpha-2 Polyclonal Antibody immunoprecipitates AMPK alpha 2 protein in IP experiments. IP Sample: 1,000 µg 293T whole cell lysate/extract A. 30 µg 293T whole cell lysate/extract B. Control with 2.5 µg of preimmune rabbit IgG C. Immunoprecipitation of AMPK alpha 2 protein by 2.5 µg of AMPK alpha-2 Polyclonal Antibody (Product # PA5-21494) 10% SDS-PAGE The immunoprecipitated AMPK alpha 2 protein was detected by AMPK alpha-2 Polyclonal Antibody (Product # PA5-21494) diluted at 1:1,000.
