Anti-SERCA2 ATPase antibody [EPR9392] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SERCA2 ATPase antibody [EPR9392] - BSA and Azide free (AB238426)

Immunohistochemical analysis of paraffin embedded Human liver tissue labelling SERCA2 ATPase with unpurified ab150435 at 1/50.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150435).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SERCA2 ATPase antibody [EPR9392] - BSA and Azide free (AB238426)

Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling SERCA2 ATPase with Purified ab150435 at 1/100 dilution (10 μg/mL). Cells were fixed in 100% Methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150435).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SERCA2 ATPase antibody [EPR9392] - BSA and Azide free (AB238426)

Immunofluorescence analysis of HeLa cells labelling SERCA2 ATPase with unpurified ab150435 at 1/100.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150435).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SERCA2 ATPase antibody [EPR9392] - BSA and Azide free (AB238426)

Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling SERCA2 ATPase with unpurified ab150435 at 1/50.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150435).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SERCA2 ATPase antibody [EPR9392] - BSA and Azide free (AB238426)

Overlay histogram showing HepG2 cells stained with unpurified ab150435 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab150435, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150435).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-SERCA2 ATPase antibody [EPR9392] - BSA and Azide free (AB238426)

Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling SERCA2 ATPase with Purified ab150435 at 1/1000 dilution (1 μg/mL) (Red). Cells were fixed with 80% Methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150435).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SERCA2 ATPase antibody [EPR9392] - BSA and Azide free (AB238426)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung carcinoma tissue sections labeling SERCA2 ATPase with Purified ab150435 at 1/50 dilution (20 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150435).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SERCA2 ATPase antibody [EPR9392] - BSA and Azide free (AB238426)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human brain tissue sections labeling SERCA2 ATPase with Purified ab150435 at 1/50 dilution (20 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150435).

• WB

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Western blot - Anti-SERCA2 ATPase antibody [EPR9392] - BSA and Azide free (AB238426)

Suggest to use non-boiled samples, as boiling process could cause membrane protein aggregates (PMID : 16023741 and PMID : 8670158).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150435).

All lanes:

Western blot - Anti-SERCA2 ATPase antibody [EPR9392] (<a href='/en-us/products/primary-antibodies/serca2-atpase-antibody-epr9392-ab150435'>ab150435</a>) at 1/5000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates boiled at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates unboiled at 20 µg

Lane 3:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates boiled at 20 µg

Lane 4:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates unboiled at 20 µg

Lane 5:

Mouse brain lysates boiled at 20 µg

Lane 6:

Mouse brain lysates unboiled at 20 µg

Lane 7:

Rat brain lysates boiled at 20 µg

Lane 8:

Rat brain lysates unboiled at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 115 kDa

Observed band size: 115 kDa,140 kDa

false