Anti-Inhibin beta E chain antibody [EPR29191-125]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Inhibin beta E chain antibody [EPR29191-125] (AB322202)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling Inhibin beta E chain with ab322202 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in human skeletal muscle.
The section was incubated with ab322202 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

Lab

Multiplex immunohistochemistry - Anti-Inhibin beta E chain antibody [EPR29191-125] (AB322202)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining Carboxypeptidase A with ab317620 at a 1/1000 dilution, ab322202 anti-Inhibin beta E chain used at 1/2000 dilution and ab52625 anti-Cytokeratin 19 used at a 1/6000 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody.

Panel A : merged staining of anti-VAP1 (green; Opal^™520), anti-Inhibin beta E chain (magenta; Opal^™690) and anti-Cytokeratin 19 (gray; Opal^™570) on human liver. Panel B : anti-VAP1 staining endothelium in human liver.
Panel C : anti-Inhibin beta E chain staining hepatocytes in human liver.
Panel D : anti-Cytokeratin 19 staining branch of bile ducts in human liver.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab317620, ab322202 and ab52625 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.

Heat mediated antigen retrieval 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-Inhibin beta E chain antibody [EPR29191-125] (AB322202)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Inhibin beta E chain with ab322202 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in human spleen.
The section was incubated with ab322202 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-Inhibin beta E chain antibody [EPR29191-125] (AB322202)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Inhibin beta E chain with ab322202 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human liver.
The section was incubated with ab322202 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-Inhibin beta E chain antibody [EPR29191-125] (AB322202)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling Inhibin beta E chain with ab322202 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in mouse skeletal muscle.
The section was incubated with ab322202 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-Inhibin beta E chain antibody [EPR29191-125] (AB322202)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Inhibin beta E chain with ab322202 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in mouse spleen.
The section was incubated with ab322202 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-Inhibin beta E chain antibody [EPR29191-125] (AB322202)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Inhibin beta E chain with ab322202 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse liver.
The section was incubated with ab322202 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-Inhibin beta E chain antibody [EPR29191-125] (AB322202)

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

Negative control : skeletal muscle, uterus (PMID : 12242034, PMID : 11932210, PMID : 9070865).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 35999217, PMID : 35896531).

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

All lanes:

Western blot - Anti-Inhibin beta E chain antibody [EPR29191-125] (ab322202) at 1/1000 dilution

Lane 1:

Human liver tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Lane 3:

Human uterus 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: 48 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Inhibin beta E chain antibody [EPR29191-125] (AB322202)

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

Negative control : skeletal muscle, spleen, ovary, uterus (PMID : 12242034, PMID : 11932210, PMID : 9070865).

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

All lanes:

Western blot - Anti-Inhibin beta E chain antibody [EPR29191-125] (ab322202) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Lane 4:

Mouse ovary tissue lysate at 20 µg

Lane 5:

Mouse uterus 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 1/100000 dilution

Observed band size: 48 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Inhibin beta E chain antibody [EPR29191-125] (AB322202)

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

This antibody does not cross-react with human Inhibin beta C chain.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Inhibin beta E chain antibody [EPR29191-125] (ab322202) at 1/1000 dilution

Lane 1:

His-tagged human Inhibin beta E chain fragment at 10 ng

Lane 2:

His-tagged human Inhibin beta C chain fragment at 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

Observed band size: 15 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Inhibin beta E chain antibody [EPR29191-125] (AB322202)

Left : Anti-Inhibin beta E chain antibody ab103167 staining at 1/1000 dilution, shown in green; Mouse anti-CANX (ab238078) loading control staining at 1/20,000 dilution, shown in magenta.

Right : Rabbit Monoclonal [EPR29191-125] to Inhibin beta E chain ab322202 staining at 1/1000 dilution, shown in green; Mouse anti-CANX (ab238078) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 43 kDa in INHBE-DDDDK HEK-293T overexpression lysate with no signal observed at this size in INHBC-DDDDK HEK-293T overexpression lysate.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution. For DDDDK-tag detection, HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (ab49763) was used at 1/1000 dilution and developed using the ECL technique.

Neither ab103167 nor ab322202 cross-reacts with Inhibin beta C chain (INHBC). In Western Blot, we recommend ab322202 as the superior alternative to ab103167 for better specificity and sensitivity.

Exposure time for DDDDK tag : 2s.

Lanes 1 - 4:

Western blot - Anti-Inhibin beta E chain antibody (<a href='/en-us/products/primary-antibodies/inhibin-beta-e-chain-antibody-ab103167'>ab103167</a>) at 1/1000 dilution

Lanes 2, 4, 6 and 8:

Western blot - HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (<a href='/en-us/products/primary-antibodies/hrp-ddddk-tag-binds-to-flag-tag-sequence-antibody-m2-ab49763'>ab49763</a>)

Lanes 5 - 8:

Western blot - Anti-Inhibin beta E chain antibody [EPR29191-125] (ab322202) at 1/1000 dilution

Lanes 1, 3, 5 and 7:

HEK-293 Mock Transfection Lysate at 20 µg

Lanes 2 and 6:

INHBE-DDDDK HEK-293T Overexpression Lysate at 20 µg

Lanes 4 and 8:

INHBC-DDDDK HEK-293T Overexpression Lysate at 20 µg

Secondary

Lanes 1 - 8:

Goat anti-Rabbit 800CW at 1/20000 dilution

Lanes 1 - 8:

Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 35 kDa

Observed band size: 43 kDa

true