Anti-Calbindin antibody [EPR22698-236]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calbindin antibody [EPR22698-236] (AB229915)

Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling Calbindin with ab229915 at 1/4000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on Purkinje cells of rat cerebellum (PMID : 17507571) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

The section was incubated with ab229915 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Calbindin antibody [EPR22698-236] (AB229915)

Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized rat cerebellum tissue labeling Calbindin with ab229915 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution (green). Positive staining on molecular layer and purkinje cells of rat cerebellum (PMID : 20130198) is observed. The nuclear counter stain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Calbindin antibody [EPR22698-236] (AB229915)

Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse cerebellum tissue labeling Calbindin with ab229915 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution (green). Positive staining on molecular layer and purkinje cells of mouse cerebellum (PMID : 20130198) is observed. The nuclear counter stain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calbindin antibody [EPR22698-236] (AB229915)

Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labeling Calbindin with ab229915 at 1/4000 dilution, followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Strong positive staining on Purkinje cells of mouse cerebellum (PMID : 17507571) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

The section was incubated with ab229915 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

• IP

Unknown

Immunoprecipitation - Anti-Calbindin antibody [EPR22698-236] (AB229915)

Calbindin was immunoprecipitated from 0.35 mg of mouse brain lysate with ab229915 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab229915 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as the secondary antibody at 1/5000 dilution.

Lane 1 : Mouse brain lysate 10 μg (Input).
Lane 2 : ab229915 IP in mouse brain lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab229915 in mouse brain lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 5 seconds.

All lanes:

Immunoprecipitation - Anti-Calbindin antibody [EPR22698-236] (ab229915)

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-Calbindin antibody [EPR22698-236] (AB229915)

Calbindin was immunoprecipitated from 0.35 mg of rat brain lysate with ab229915 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab229915 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as the secondary antibody at 1/5000 dilution.

Lane 1 : Rat brain lysate 10 μg (Input).
Lane 2 : ab229915 IP in rat brain lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab229915 in rat brain lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-Calbindin antibody [EPR22698-236] (ab229915)

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

• WB

Unknown

Western blot - Anti-Calbindin antibody [EPR22698-236] (AB229915)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times.

Lanes 1 & 2 : 15 seconds; Lanes 3 & 4 : 3 minutes; Lane 5 : 48 seconds.

Negative control : NIH/3T3, C6 (lanes 2 & 4, PMID : 8370070, 9889325).

All lanes:

Western blot - Anti-Calbindin antibody [EPR22698-236] (ab229915) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 20 µg

Lane 2:

NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 20 µg

Lane 3:

Rat brain lysate at 20 µg

Lane 4:

C6 (rat glial tumor glial cell line) whole cell lysate at 20 µg

Lane 5:

Mouse cerebellum lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 30 kDa

Observed band size: 30 kDa

false