Anti-Caldesmon/CDM antibody [SP226] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Caldesmon/CDM antibody [SP226] - BSA and Azide free (AB238782)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) labeling Caldesmon/CDM with purified ab183339 at 1/230 dilution (10.05μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotypecontrol - Rabbit monoclonal IgG (ab172730) (black). Unlableled control - Unlabelled cells (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183339).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Caldesmon/CDM antibody [SP226] - BSA and Azide free (AB238782)

Intracellular flow cytometric analysis of HeLa cells labeling Caldesmon/CDM with ab183339 at 1/400 dilution (green) compared to a negative control Rabbit IgG (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183339).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [SP226] - BSA and Azide free (AB238782)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded human colon tissue labeling Caldesmon/CDM with ab183339 at 1/100 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab183339).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [SP226] - BSA and Azide free (AB238782)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling Caldesmon/CDM with ab183339 at 1/100 dilution (23.11 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on smooth muscle in human colon, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab183339 for 10 mins at room temperature. This image was generated using ab183339, the same clone, but with a different buffer formulation.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [SP226] - BSA and Azide free (AB238782)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse colon tissue sections labeling Caldesmon/CDM with ab183339 at 1/100 dilution (23.11 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on smooth muscle in mouse colon, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab183339 for 10 mins at room temperature. This image was generated using ab183339, the same clone, but with a different buffer formulation.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [SP226] - BSA and Azide free (AB238782)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat colon tissue sections labeling Caldesmon/CDM with ab183339 at 1/100 dilution (23.11 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on smooth muscle in rat colon, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab183339 for 10 mins at room temperature. This image was generated using ab183339, the same clone, but with a different buffer formulation.

• WB

Lab

Western blot - Anti-Caldesmon/CDM antibody [SP226] - BSA and Azide free (AB238782)

This data was developed using the same antibody clone in a different buffer formulation (ab183339).

Lanes 1-4 : Merged signal (red and green). Green - ab183339 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.

ab183339 Anti-Caldesmon/CDM antibody [SP226] was shown to specifically react with Caldesmon/CDM in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265026 (knockout cell lysate ab257375) was used. Wild-type and Caldesmon/CDM knockout samples were subjected to SDS-PAGE. ab183339 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Caldesmon/CDM antibody [SP226] (<a href='/ko/products/primary-antibodies/caldesmon-cdm-antibody-sp226-ab183339'>ab183339</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CALD1 (Caldesmon/CDM) knockout HeLa cell lysate (<a href='/ko/products/cell-lysates/human-cald1-caldesmon-cdm-knockout-hela-cell-lysate-ab257375'>ab257375</a>) at 20 µg

Lane 3:

NIH/3T3 cell lysate at 20 µg

Lane 4:

HEK-293 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 93 kDa

Observed band size: 75 kDa

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