Anti-Cystatin F antibody [EPR29573-88]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin F antibody [EPR29573-88] (AB325174)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Cystatin F with ab325174 at 1/2000 (0.252 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on mouse cerebrum. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin F antibody [EPR29573-88] (AB325174)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a Mouse CST7 expression vector containing a His tag and (B) HEK-293T transfected with empty vector containing a His tag tissue labeling Cystatin F with ab325174 at 1/5000 (0.101 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on HEK-293T transfected with a His-tagged Mouse CST7 construct (image A) and no staining on (B) HEK-293T transfected with empty vector containing a His tag. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin F antibody [EPR29573-88] (AB325174)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Cystatin F with ab325174 at 1/2000 (0.252 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on rat cerebrum. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin F antibody [EPR29573-88] (AB325174)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling Cystatin F with ab325174 at 1/2000 (0.252 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on rat cardiac muscle. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin F antibody [EPR29573-88] (AB325174)

Immunohistochemical analysis of paraffin-embedded Rat bone marrow tissue labeling Cystatin F with ab325174 at 1/2000 (0.252 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat bone marrow. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin F antibody [EPR29573-88] (AB325174)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling Cystatin F with ab325174 at 1/2000 (0.252 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on mouse skeletal muscle. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin F antibody [EPR29573-88] (AB325174)

Immunohistochemical analysis of paraffin-embedded Mouse bone marrow tissue labeling Cystatin F with ab325174 at 1/2000 (0.252 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse bone marrow. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin F antibody [EPR29573-88] (AB325174)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Cystatin F with ab325174 at 1/2000 (0.252 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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

Lab

Western blot - Anti-Cystatin F antibody [EPR29573-88] (AB325174)

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

Low expression : brain(PMID : 9733783)

The loading buffer in the lysate contains DTT.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Cystatin F antibody [EPR29573-88] (ab325174) at 1/1000 dilution

Lane 1:

Rat bone marrow lysate at 20 µg

Lane 2:

Rat brain tissue 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

Observed band size: 15-18 kDa,36 kDa

false

Exposure time: 48s

• WB

Lab

Western blot - Anti-Cystatin F antibody [EPR29573-88] (AB325174)

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

Low expression : 4T1, F9

The loading buffer in the lysate contains DTT.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) 1 : 100000 (15Kda).

All lanes:

Western blot - Anti-Cystatin F antibody [EPR29573-88] (ab325174) at 1/1000 dilution

Lane 1:

P815 (mouse mastocytoma mast Cell ) whole cell lysate at 20 µg

Lane 2:

4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

F9 (mouse embryonal carcinoma epithelial cell) whole cell 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

Observed band size: 15-18 kDa,15 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-Cystatin F antibody [EPR29573-88] (AB325174)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 18256700).

CST7 is a 15KD glycoprotein and the 30-37KD band represents the homodimer.

DTT is used to break dimers

All lanes:

Western blot - Anti-Cystatin F antibody [EPR29573-88] (ab325174) at 1/1000 dilution

Lane 1:

Mouse bone marrow lysate without DTT at 20 µg

Lane 2:

Mouse bone marrow lysate with DTT 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

Observed band size: 15 kDa,30-37 kDa

false

Exposure time: 48s

• WB

Lab

Western blot - Anti-Cystatin F antibody [EPR29573-88] (AB325174)

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

Low expression : brain, skeletal muscle(PMID : 9733783)

The loading buffer in the lysate contains DTT

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Cystatin F antibody [EPR29573-88] (ab325174) at 1/1000 dilution

Lane 1:

Mouse bone marrow lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 5:

EL4 (mouse lymphoma T lymphocyte) whole cell 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

Observed band size: 15-18 kDa,36 kDa

false

Exposure time: 48s