Anti-SFTPA1 antibody [EPR29098-90R]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SFTPA1 antibody [EPR29098-90R] (AB325321)

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling SFTPA1 with ab325321 at 1/1000 (0.512 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on type II pneumocytes in human lung (PMID : 32049827).
The primary antibody was incubated 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).

Heat mediated antigen retrieval was performed 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-SFTPA1 antibody [EPR29098-90R] (AB325321)

Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling SFTPA1 with ab325321 at 1/1000 (0.512 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human lung carcinoma.
The primary antibody was incubated 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).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• mIHC

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Multiplex immunohistochemistry - Anti-SFTPA1 antibody [EPR29098-90R] (AB325321)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human lung tissue staining SFTPA1 with ab325321 at a 1 : 1000 (0.512 ug/ml) dilution, ab213363 anti-CD68 used at 1 : 500 (1.314 ug/ml) dilution and ab239904 anti-Aquaporin 5 used at a 1 : 500 (2.0 ug/ml) dilution.

Panel A : merged staining of anti-SFTPA1 (green; Opal™520), anti-CD68 (magenta; Opal™690) and anti-Aquaporin 5 (gray; Opal™570) on human lung.
Panel B : anti-SFTPA1 staining type 2 pneumonocytes in human lung.
Panel C : anti-CD68 staining macrophages in human lung.
Panel D : anti-Aquaporin 5 staining epithelium and type 1 pneumonocytes in human lung.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325324, ab213363 and ab239904 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.

Nuclear counter stain with DAPI.

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

• WB

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Western blot - Anti-SFTPA1 antibody [EPR29098-90R] (AB325321)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : cerebellum, spleen, liver (PMID : 34181042).

The identity of the lower MW band at approximately 17 kDa (in lane 2) is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-SFTPA1 antibody [EPR29098-90R] (ab325321) at 1/1000 dilution

Lane 1:

Human lung tissue lysate at 20 µg

Lane 2:

Human cerebellum tissue lysate at 20 µg

Lane 3:

Human spleen tissue lysate at 20 µg

Lane 4:

Human liver tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 36 kDa

false

Exposure time: 48s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SFTPA1 antibody [EPR29098-90R] (AB325321)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling SFTPA1 with ab325321 at 1/1000 (0.512 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on human kidney (HPA).
The primary antibody was incubated 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).

Heat mediated antigen retrieval was performed 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-SFTPA1 antibody [EPR29098-90R] (AB325321)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling SFTPA1 with ab325321 at 1/1000 (0.512 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on human spleen (HPA).
The primary antibody was incubated 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).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-SFTPA1 antibody [EPR29098-90R] (AB325321)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-reacts with overexpressed human SFTPA2.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-SFTPA1 antibody [EPR29098-90R] (ab325321) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate at 5 µg

Lane 2:

293T cells transfected with a human SFTPA1 expression vector containing a myc-His-tag® whole cell lysate at 5 µg

Lane 3:

293T cells transfected with a human SFTPA2 expression vector containing a myc-His-tag® whole cell lysate at 5 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 1/100000 dilution

Observed band size: 35 kDa,124 kDa

false

Exposure time: 10s