Anti-CaSR antibody [EPR24050-59]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CaSR antibody [EPR24050-59] (AB259846)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling CaSR with ab259846 at 1/5000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining in human pancreatic islet and ducts. The section was incubated with ab259846 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CaSR antibody [EPR24050-59] (AB259846)

Immunohistochemical analysis of paraffin-embedded Human parathyroid gland tissue labeling CaSR with ab259846 at 1/5000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining in human parathyroid gland. The section was incubated with ab259846 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CaSR antibody [EPR24050-59] (AB259846)

Fluorescence multiplex immunohistochemical analysis of the Human parathyroid gland (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-CaSR (ab259846, magenta; Opal™690), anti-Cytochrome C (ab247438, green; Opal™520) and anti-FABP4 (ab92501, red; Opal™570) on human parathyroid gland. Panel B : anti-CaSR stained on parathyroid chief cells. Panel C : anti-Cytochrome C stained on parathyroid oxyphil cells. Panel D : anti-FABP4 stained on adipocytes. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The section was incubated in three rounds of staining : in the order of ab259846 at 1/5000 dilution (0.103 μg/ml), ab247438 at 1/5000 dilution (0.195 μg/ml), and ab92501 at 1/10000 dilution (0.047 μg/ml) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CaSR antibody [EPR24050-59] (AB259846)

Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue labeling CaSR with ab259846 at 1/5000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining in human cerebellum. The section was incubated with ab259846 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

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

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CaSR antibody [EPR24050-59] (AB259846)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat kidney tissue labeling CaSR with ab259846 at 1/50 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on rat kidney is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CaSR antibody [EPR24050-59] (AB259846)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse thyroid staining of Thyroid Peroxidase/TPO with ab278525 at a 1/5000 (0.109 μg/ml) dilution, PAX8 with ab239363 at 1/2000 (0.254 μg/ml) dilution, and CaSR with ab259846 at 1/5000 (0.103 μg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Thyroid peroxidase (green; Opal™ 520), anti-PAX8 (magenta; Opal™ 690) and anti-CaSR (gray; Opal™ 570) on mouse thyroid.
Panel B : anti-Thyroid peroxidase showed cytoplasmic and membranous staining on mouse thyroid.
Panel C : anti-PAX8 showed nucleus staining on mouse thyroid.
Panel D : anti-CaSR staining parafollicular cells.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab278525, ab239363 and ab259846 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CaSR antibody [EPR24050-59] (AB259846)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse kidney tissue labeling CaSR with ab259846 at 1/50 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse kidney is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CaSR antibody [EPR24050-59] (AB259846)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling CaSR with ab259846 at 1/5000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in mouse kidney. The section was incubated with ab259846 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CaSR antibody [EPR24050-59] (AB259846)

Immunohistochemical analysis of paraffin-embedded Rat parathyroid gland tissue labeling CaSR with ab259846 at 1/5000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining in rat parathyroid gland. The section was incubated with ab259846 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• WB

Lab

Western blot - Anti-CaSR antibody [EPR24050-59] (AB259846)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as a GAPDH loading control. Negative control : skeletal muscle. The molecular weight observed is consistent with literatures (PMID : 9722601, PMID : 3360311). Samples are non-boiled as boiling may cause protein aggregation and were run on a Bis-Tris gel. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CaSR antibody [EPR24050-59] (ab259846) at 1/1000 dilution

Lane 1:

Human kidney tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 5:

Rat kidney tissue lysate at 20 µg

Lane 6:

Rat skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 121 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-CaSR antibody [EPR24050-59] (AB259846)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab181602 and ab179475 were used as loading controls. The molecular weight observed is consistent with literatures (PMID : 9722601, PMID : 3360311). Samples are non-boiled as boiling may cause protein aggregation and were run on a Bis-Tris gel. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution and anti- Integrin alpha V antibody (ab179475) loading control staining at 1/5000 dilution.

All lanes:

Western blot - Anti-CaSR antibody [EPR24050-59] (ab259846) at 1/1000 dilution

Lane 1:

Mouse kidney membrane tissue lysate at 10 µg

Lane 2:

Mouse kidney membrane control tissue lysate at 10 µg

Lane 3:

Rat kidney membrane tissue lysate at 10 µg

Lane 4:

Rat skeletal muscle tissue lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

false

Exposure time: 10s