Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (AB215275)

This data was developed using ab32330, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human leiomyoma tissue sections labeling Caldesmon/CDM with purified ab32330 at 1/100 dilution (1.21 μg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (AB215275)

This data was developed using ab32330, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomach tissue sections labeling Caldesmon/CDM with purified ab32330 at 1/100 dilution (1.21 μg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (AB215275)

This data was developed using ab32330, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling Caldesmon/CDM with purified ab32330 at 1/100 dilution (1.21 μg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (AB215275)

This data was developed using ab32330, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling Caldesmon/CDM with purified ab32330 at 1/50 dilution (2.4 μg/mL). Cells were fixed in 100% Methanol and permeabilized with 0.1% TritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (AB215275)

This data was developed using ab32330, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labelling Caldesmon/CDM with purified ab32330 at 1/20 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IP

Lab

Immunoprecipitation - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (AB215275)

This data was developed using ab32330, the same antibody clone in a different buffer formulation.
Purified ab32330 at 1/20 dilution (0.6μg) immunoprecipitating Caldesmon/CDM in NIH/3T3 whole cell lysate.
Lane 1 (input) : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate.
Lane 2 (+) : ab32330 + NIH/3T3 whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab32330 in NIH/3T3 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Caldesmon/CDM antibody [E89] (<a href='/en-us/products/primary-antibodies/caldesmon-cdm-antibody-e89-ab32330'>ab32330</a>)

Predicted band size: 93 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (AB215275)

This data was developed using the same antibody clone in a different buffer formulation (ab32330).

Lanes 1-4 : Merged signal (red and green). Green - ab32330 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.

ab32330 Anti-Caldesmon/CDM antibody [E89] 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. ab32330 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 [E89] (<a href='/en-us/products/primary-antibodies/caldesmon-cdm-antibody-e89-ab32330'>ab32330</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CALD1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CALD1 (Caldesmon/CDM) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cald1-caldesmon-cdm-knockout-hela-cell-line-ab265026'>ab265026</a>)

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='/en-us/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

false

• WB

Unknown

Western blot - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (AB215275)

All lanes:

Western blot - Anti-Caldesmon/CDM antibody [E89] (<a href='/en-us/products/primary-antibodies/caldesmon-cdm-antibody-e89-ab32330'>ab32330</a>) at 1/10000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 15 µg

Lane 3:

Rat liver lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 93 kDa

false