Anti-Proenkephalin-B antibody [EPR24508-78]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proenkephalin-B antibody [EPR24508-78] (AB283501)

Immunohistochemical analysis of paraffin-embedded Rat hippocampus tissue labelling ProDynorphin with ab283501 at 1/4000 (0.137 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat hippocampus (PMID : 16966485). The section was incubated with ab283501 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-Proenkephalin-B antibody [EPR24508-78] (AB283501)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labelling ProDynorphin with ab283501 at 1/4000 (0.137 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : no staining on rat liver (PMID 23649822). The section was incubated with ab283501 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Proenkephalin-B antibody [EPR24508-78] (AB283501)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver tissue labeling ProDynorphin with ab283501 at 1/100 (5.47 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 2 ug/ml dilution (Green). No staining on rat liver.Negative control : liver (PMID : 3864164) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 2 ug/ml dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proenkephalin-B antibody [EPR24508-78] (AB283501)

Immunohistochemical analysis of paraffin-embedded Rat pituitary tissue labelling ProDynorphin with ab283501 at 1/4000 (0.137 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat pituitary (PMID : 16966485). The section was incubated with ab283501 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IP

Supplier Data

Immunoprecipitation - Anti-Proenkephalin-B antibody [EPR24508-78] (AB283501)

ProDynorphin was immunoprecipitated from 0.35 mg Rat pituitary tissue lysate with ab283501 at 1/30 dilution (2 ug in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab283501 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Rat pituitary tissue lysate 10 ug

Lane 2 : ab283501 IP in Rat pituitary tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283501 in rat pituitary tissue lysate

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

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-Proenkephalin-B antibody [EPR24508-78] (ab283501)

Predicted band size: 28 kDa

Observed band size: 18 kDa,23 kDa,28 kDa

false

• WB

Lab

Western blot - Anti-Proenkephalin-B antibody [EPR24508-78] (AB283501)

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

The observed molecular weight is consistent with the literature (PMID : 9712897).

Negative control : Rat liver (PMID : 23649822).

Exposure time : 26 seconds.

All lanes:

Western blot - Anti-Proenkephalin-B antibody [EPR24508-78] (ab283501) at 1/1000 dilution

Lane 1:

Rat pituitary tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 40 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: 28 kDa

Observed band size: 18 kDa,23 kDa,28 kDa

false

• WB

Lab

Western blot - Anti-Proenkephalin-B antibody [EPR24508-78] (AB283501)

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

Exposure time : 1 second.

All lanes:

Western blot - Anti-Proenkephalin-B antibody [EPR24508-78] (ab283501) at 1/1000 dilution

Lane 1:

LIF/His-rat ProDynorphin full-length protein at 0.01 µg

Lane 2:

His-tagged rat ProDynorphin pro-peptide at 0.01 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: 28 kDa

Observed band size: 18 kDa,64 kDa

false

• Dot

Supplier Data

Dot Blot - Anti-Proenkephalin-B antibody [EPR24508-78] (AB283501)

Dot blot analysis of ProDynorphin using ab283501 at 1/1000 (0.583 ug/ml) diultion followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution.

Exposure time : 3 minutes.

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