Anti-Calreticulin antibody [EPR3924] - BSA and Azide free

• IHC

Lab

Immunohistochemistry - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

Immunohistochemical analysis of formalin fixed paraffin embedded human kidney labelling Calreticulin with ab92516 at a concentration of 0.004 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5 for 32 mins.

ab92516 anti-Calreticulin [EPR3924] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

Formalin-fixed, paraffin-embedded normal human stomach tissue stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling Calreticulin with ab92516, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining human breast carcinoma.The section was incubated with ab92516 for 30 mins at room temperature. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

ab92516 staining Calreticulin in wild-type HAP1 cells (top panel) and CALR knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92516 at 1/500 and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor^® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Alexa Fluor^® 488 (ab196158) and Alexa Fluor^® 647 (ab196159) conjugated versions are available for this clone.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

ab92516 showing negative staining in Normal human heart tissue.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

Overlay histogram showing HAP1 wildtype (green line) and HAP1-CALR knockout cells (red line) stained with ab92516. The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab92516, 1μg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) preadsorbed (ab150081) at 1/2000 dilution for 30 min at 22°C. A rabbit IgG isotype control antibody (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-CALR knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

Alexa Fluor^® 488 (ab196158) and Alexa Fluor^® 647 (ab196159) conjugated versions are available for this clone.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

Formalin-fixed, paraffin-embedded Papillary carcinoma of human thyroid gland tissue.stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

Immunocytochemistry/Immunofluorescence analysis of HT-29 (human colorectal adenocarcinoma) labelling Calreticulin with purified ab92516 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (ab150077). Nuclei counterstained with DAPI (blue).

Control : PBS only

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

ab92516, at 1/250 dilution, staining Calreticulin in paraffin embedded Human kidney tissue by Immunohistochemistry.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

Formalin-fixed, paraffin-embedded normal human liver tissue stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

Overlay histogram showing HeLa cells stained with ab92516 (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 (ab92516, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

Formalin-fixed, paraffin-embedded normal human placenta tissue stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

Formalin-fixed, paraffin-embedded normal human colon tissue stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling Calreticulin with ab92516, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on rat lung.The section was incubated with ab92516 for 30 mins at room temperature. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (AB271865)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Calreticulin with ab92516, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on mouse liver.The section was incubated with ab92516 for 30 mins at room temperature. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).