Anti-SERCA2 ATPase 抗体

• WB

Supplier Data

Western blot - Anti-SERCA2 ATPase antibody (AB3625)

All lanes:

Western blot - Anti-SERCA2 ATPase antibody (ab3625) at 0.1 µg/mL

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 50 µg

Lane 2:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 50 µg

Lane 3:

NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 50 µg

Predicted band size: 115 kDa

true

Exposure time: 30s

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SERCA2 ATPase antibody (AB3625)

ab3625 staining human heart. Staining is localised to the cytoplasm/sarcoplasm.
Left panel : with primary antibody at 1 ug/ml. Right panel : isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the DAKO 3-in-1 antigen retrieval buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SERCA2 ATPase antibody (AB3625)

ICC/IF image of ab3625 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3625, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• WB

Supplier Data

Western blot - Anti-SERCA2 ATPase antibody (AB3625)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Anti-SERCA2 ATPase antibody (ab320749, Left) staining at 1/1000 dilution showed similar expression profile/ molecular weight with Anti-SERCA2 ATPase antibody (ab3625, Right) at 1/1000 dilution. Suggest to use non-boiled samples, as boiling process could cause membrane protein aggregates (PMID : 16023741 and PMID : 8670158).

All lanes:

Western blot - Anti-SERCA2 ATPase antibody [RM1176] (<a href='/products/primary-antibodies/serca2-atpase-antibody-rm1176-ab320749'>ab320749</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate boiled at 20 µg

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate unboiled at 20 µg

Lane 3:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate boiled at 20 µg

Lane 4:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate unboiled at 20 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate boiled at 20 µg

Lane 6:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate unboiled at 20 µg

Lane 7:

H9c2 (2-1) (rat cardiomyoblast) whole cell lysate boiled at 20 µg

Lane 8:

H9c2 (2-1) (rat cardiomyoblast) whole cell lysate unboiled at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 115 kDa,140 kDa

false

Exposure time: 26s

• IP

Supplier Data

Immunoprecipitation - Anti-SERCA2 ATPase antibody (AB3625)

SERCA2 ATPase was immunoprecipitated from HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (1 mg for IP, 20% of IP loaded) with ab3625 at 6 μg/mg lysate. Western blot was performed from the immunoprecipitate using ab3625 at 1 μg/ml.

Lane 1 : ab3625 IP in HEK-293T whole cell lysate.

Lane 3 : ab3625 IP in HEK-293T whole cell lysate.

Lane 2 : Control IgG IP in HEK-293T whole cell lysate.

Detection : Chemiluminescence with exposure time of 3 minutes.

All lanes:

Immunoprecipitation - Anti-SERCA2 ATPase antibody (ab3625)

Predicted band size: 115 kDa

true

• WB

Unknown

Western blot - Anti-SERCA2 ATPase antibody (AB3625)

Sample : Homogenate from Pig (lanes 1 and 2) or Rabbit (lane 4) aorta or lysate from cultured Rat aortic smooth muscle cells (lane 3)

Affinity purified Rabbit anti-SERCA2 (ab3625) (lanes 2, 3 and IP for lane 4) or control (ctrl) monoclonal anti-SERCA2 (lanes 1 and WB for lane 4).

Dilutions : 1 : 1,000 (lanes 1, 2 and 4) or 1 : 2,500 (lane 3)

All lanes:

Western blot - Anti-SERCA2 ATPase antibody (ab3625)

Predicted band size: 115 kDa

false