Anti-Surfactant protein D/SP-D antibody [EPR21928-209]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Surfactant protein D/SP-D antibody [EPR21928-209] (AB220423)

Immunohistochemical analysis of paraffin-embedded human lung tissue labeling Surfactant protein D/SP-D with ab220423 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on alveolar type II cells of human lung (PMID : 10820266) is observed. Counter stained with Hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Surfactant protein D/SP-D antibody [EPR21928-209] (AB220423)

Negative control : No staining on human skeletal muscle (PMID : 10640760) is observed

Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling Surfactant protein D/SP-D with ab220423 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Surfactant protein D/SP-D antibody [EPR21928-209] (AB220423)

Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded Human lung tissue. Panel A : Merged staining of anti-Podoplanin / gp36 (magenta; Opal™690), anti-Surfactant protein D/SP-D (green; Opal™520) and anti-Sialoadhesin/CD169 (red; Opal™570) on human lung. Panel B : Anti-Podoplanin / gp36 stained on alveolar type I cells. Panel C : Anti-Surfactant protein D/SP-D stained on alveolar type II cells. Panel D : Anti-Sialoadhesin/CD169 stained on macrophages. The section was incubated in three rounds of staining : in the order of ab236529 and ab234257 for 30 mins, then ab245735 for 10 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 solution 2) for 20 mins. Counterstained with DAPI.

• WB

Supplier Data

Western blot - Anti-Surfactant protein D/SP-D antibody [EPR21928-209] (AB220423)

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular mass observed is consistent with what has been described in the literature (PMID : 9751757).

All lanes:

Western blot - Anti-Surfactant protein D/SP-D antibody [EPR21928-209] (ab220423) at 1/1000 dilution

All lanes:

Human lung lysate at 20 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: 38 kDa

Observed band size: 43 kDa

true

Exposure time: 3min

• IP

Supplier Data

Immunoprecipitation - Anti-Surfactant protein D/SP-D antibody [EPR21928-209] (AB220423)

Surfactant protein D/SP-D was immunoprecipitated from 0.35 mg human fetal lung lysate with ab220423 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220423 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1 : Human fetal lung lysate 10 μg (Input).
Lane 2 : ab220423 IP in human fetal lung lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab220423 in human fetal lung lysate.
Blocking/Dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 minutes.

The molecular mass observed is consistent with what has been described in the literature (PMID : 9751757).

All lanes:

Immunoprecipitation - Anti-Surfactant protein D/SP-D antibody [EPR21928-209] (ab220423)

Predicted band size: 38 kDa

Observed band size: 43 kDa

false

• WB

Supplier Data

Western blot - Anti-Surfactant protein D/SP-D antibody [EPR21928-209] (AB220423)

Blocking/Dilution buffer : 5% NFDM/TBST.

Recombinant protein was obtained from a mammalian expression system.

All lanes:

Western blot - Anti-Surfactant protein D/SP-D antibody [EPR21928-209] (ab220423) at 1/1000 dilution

All lanes:

Human Surfactant protein D/SP-D recombinant protein (aa 21-375), 10 ng

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 1/100000 dilution

Predicted band size: 38 kDa

false

Exposure time: 32s