- MA5-33000 - Invitrogen Antibodies HSPA2 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HSPA2 in Lane 1: human HeLa whole cell lysate, Lane 2: human MDA-MB-231 whole cell lysate, Lane 3: human COLO-320 whole cell lysate, Lane 4: human PANC-1 whole cell lysate. Electrophoresis was performed with 5-20% SDS-PAGE gel (70V, Stacking gel; 90V Resolving gel, Time: 2-3 hours), transferred to a nitrocellulose membrane and blocked using 5% Non-fat Milk/TBS (1.5 hrs at room temperature). Samples were incubated with HSPA2 monoclonal antibody (Product # MA5-33000) using a 0.5 µg/mL dilution, followed by a goat anti-mouse IgG-HRP at a dilution of 1:10,000, and developed with enhanced chemiluminescence (ECL).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HSPA2 in Lane 1: human HeLa whole cell lysate, Lane 2: human MDA-MB-231 whole cell lysate, Lane 3: human COLO-320 whole cell lysate, Lane 4: human PANC-1 whole cell lysate. Electrophoresis was performed with 5-20% SDS-PAGE gel (70V, Stacking gel; 90V Resolving gel, Time: 2-3 hours), transferred to a nitrocellulose membrane and blocked using 5% Non-fat Milk/TBS (1.5 hrs at room temperature). Samples were incubated with HSPA2 monoclonal antibody (Product # MA5-33000) using a 0.5 µg/mL dilution, followed by a goat anti-mouse IgG-HRP at a dilution of 1:10,000, and developed with enhanced chemiluminescence (ECL).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HSPA2 in, Lane 1: rat lung tissue lysates, Lane 2: rat liver tissue lysates, Lane 3: rat kidney tissue lysates, Lane 4: rat testicular tissue lysates, Lane 5: mouse lung tissue lysates, Lane 6: mouse liver tissue lysates, Lane 7: mouse kidney tissue lysates, Lane 8: mouse testicular tissue lysates, Lane 9: mouse RAW246. 7 whole cell lysates, . Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. The membrane was blocked with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with HSPA2 Monoclonal Antibody (4A4) (Product # MA5-33000) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0. 1% Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for HSPA2 at approximately 70KD. The expected band size for HSPA2 is at 70KD.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry analysis of HSPA2 using anti-HSPA2 antibody (Product # MA5-33000) . HSPA2 was detected in a section of PC-3 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 2μg/mL rabbit anti-HSPA2 antibody (Product # MA5-33000) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of HSPA2 in paraffin-embedded human lung cancer tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with HSPA2 monoclonal antibody (Product # MA5-33000) at a 2 µg/mL dilution, followed by biotinylated goat anti-mouse IgG, and developed with Strepavidin-Biotin-Complex and DAB.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow Cytometry of HSPA2 in PC-3 cells (blue line), isotype control mouse IgG (green line), and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with HSPA2 Monoclonal Antibody (4A4) (Product # MA5-33000) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.

MA5-33000