Anti-POMC antibody [EPR22534-165]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-POMC antibody [EPR22534-165] (AB254257)

Immunohistochemical analysis of paraffin-embedded human pituitary tissue labeling POMC with ab254257 at 1/4000 dilution at room temperature for 30 mins, followed by ready to use rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human pituitary is observed (PMID : 21622576). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was performed. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-POMC antibody [EPR22534-165] (AB254257)

Immunohistochemical analysis of 0.2% Triton X-100 permeabilized, 4% PFA fixed mouse pituitary tissue labeling POMC with ab254257 at 1/500 dilution, followed by AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. DAPI was used as nuclear counterstain. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) was performed. Positive staining on mouse pituitary (PMID : 18601691).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-POMC antibody [EPR22534-165] (AB254257)

Negative control : No staining on mouse spleen (PMID : 10395956)

Immunohistochemical analysis of paraffin-embedded mouse pituitary tissue labeling POMC with ab254257 at 1/4000 dilution at room temperature for 30 mins, followed by ready to use rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was performed. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-POMC antibody [EPR22534-165] (AB254257)

Immunohistochemical analysis of paraffin-embedded rat pituitary tissue labeling POMC with ab254257 at 1/4000 dilution at room temperature for 30 mins, followed by ready to use rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat pituitary is observed (PMID : 21622576). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was performed. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-POMC antibody [EPR22534-165] (AB254257)

Immunohistochemical analysis of paraffin-embedded mouse pituitary tissue labeling POMC with ab254257 at 1/4000 dilution at room temperature for 30 mins, followed by ready to use rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse pituitary is observed (PMID : 21622576). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was performed. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-POMC antibody [EPR22534-165] (AB254257)

Immunohistochemical analysis of 0.2% Triton X-100 permeabilized, 4% PFA fixed rat pituitary tissue labeling POMC with ab254257 at 1/500 dilution, followed by AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. DAPI was used as nuclear counterstain. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) was performed. Positive staining on rat pituitary (PMID : 18601691).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-POMC antibody [EPR22534-165] (AB254257)

Negative control : No stain on liver (PMID : 14578285)

Immunohistochemical analysis of 0.2% Triton X-100 permeabilized, 4% PFA fixed rat liver tissue labeling POMC with ab254257 at 1/100 dilution, followed by AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. DAPI was used as nuclear counterstain. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) was performed. No staining on rat liver.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-POMC antibody [EPR22534-165] (AB254257)

Immunocytochemical analysis of 100% methanol fixed, 0.1% TritonX-100 permeabilised AtT-20 cells labeling POMC with ab254257 at 1/100 dilution. ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as a secondary antibody at 1/1000 dilution. Counterstained with ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution. Nuclear staining : DAPI.

Confocal image showing cytoplasmic staining in AtT-20 cell line

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-POMC antibody [EPR22534-165] (AB254257)

Negative control : No stain on liver (PMID : 14578285)

Immunohistochemical analysis of 0.2% Triton X-100 permeabilized, 4% PFA fixed mouse liver tissue labeling POMC with ab254257 at 1/100 dilution, followed by AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. DAPI was used as nuclear counterstain. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) was performed. No staining on mouse liver.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution

• WB

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Western blot - Anti-POMC antibody [EPR22534-165] (AB254257)

Exposure time :
Lanes 1-2 : 3 seconds;
Lanes 3-4 : 10 seconds;
Lanes 5-6 : 32 seconds

Blocking buffer and concentration : 5% NFDM/TBST

Negative control : liver (lanes 2, 4 and 6, PMID : 10395956)

POMC is a glycoprotein and can be processed into 16kDa N-POMC. The molecular weight observed is consistent with what has been described in the literature (PMID 1814247, PMID 26880796, PMID 20639461).

Lanes 1 - 4:

Western blot - Anti-POMC antibody [EPR22534-165] (ab254257) at 1/5000 dilution

Lanes 5 - 6:

Western blot - Anti-POMC antibody [EPR22534-165] (ab254257) at 1/1000 dilution

Lane 1:

Mouse pituitary lysate at 20 µg

Lane 2:

Mouse liver lysate at 20 µg

Lane 3:

Human pituitary lysate at 20 µg

Lane 4:

Human liver lysate at 20 µg

Lane 5:

Rat pituitary lysate at 20 µg

Lane 6:

Rat liver 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

Predicted band size: 29 kDa

Observed band size: 16 kDa,35 kDa

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