- PA5-18205 - Invitrogen Antibodies ATP6AP2 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of ATP6IP2 using ATP6IP2 Polyclonal Antibody (Product # PA5-18205) (0.5 µg/mL) in staining of Human Cerebellum (A) and Heart (B) lysate (35 µg protein in RIPA buffer). Detected by chemiluminescence.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ATP6IP2 using ATP6IP2 Polyclonal Antibody (Product # PA5-18205) (0.5 µg/mL) in staining of Rat Heart lysate (35 µg protein in RIPA buffer). Detected by chemiluminescence.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of ATP6IP2 was achieved by transfecting MCF-7 with ATP6IP2 specific siRNAs (Silencer® select Product # s19791, s19790). Western blot analysis (Fig. a) was performed using whole cell extracts from the ATP6IP2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Arp3 Polyclonal Antibody (Product # PA5-18205, 2 µg/mL) and Rabbit anti-Goat IgG Heavy Chain Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 1:4,000 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 ATP6IP2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-ATP6IP2 Polyclonal Antibody (Product # PA5-18205) and a ~37 kD band was observed across the panel tested. An uncharacterized band (*) at ~80 kDa was also detected in in all the cell lines. Whole cell extracts (30 µg lysate) of MCF-7 (Lane 1), T-47D (Lane 2), PC-3 (Lane 3), SiHa (Lane 4), Panc-1 (Lane 5) and HCT 116 (Lane 6) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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 (2 µg/mL) and detected by chemiluminescence with Rabbit anti-Goat IgG Heavy Chain Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 1:4,000 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)
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Experimental details: Flow cytometric analysis of ATP6IP2 in HeLa cells using a polyclonal antibody (Product #PA5-18205). HeLa cells (blue line) were paraformaldehyde fixed and permeabilized with 0.5% Triton. The primary antibody was incubated for one hour (10 µg/mL) followed by an Alexa Fluor 488 secondary antibody (2 µg/mL). IgG control: Unimmunized goat IgG (black line) followed by an Alexa Fluor 488 secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow Cytometry analysis of ATP6IP2 using ATP6IP2 Polyclonal Antibody (Product # PA5-18205) in paraformaldehyde fixed HeLa cells (blue line), permeabilized with 0.5% Triton. Primary incubation 1hr (10 µg/mL) followed by Alexa Fluor 488 secondary antibody (2 µg/mL). IgG control: Unimmunized goat IgG (black line) followed by Alexa Fluor 488 secondary antibody.

PA5-18205