Anti-REG1B antibody [EPR28864-4]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-REG1B antibody [EPR28864-4] (AB323315)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human small intestine tissue staining MUC17 with ab316971 at 1/500 dilution, ab272692 anti-MUC2 used at 1/2000 dilution and ab323315 anti-REG1B used at a 1/5000 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-MUC17 (green; Opal^™570), anti-MUC2 (magenta; Opal^™690) and anti-REG1B (grey; Opal^™520) on human small intestine.
Panel B : anti-MUC17 staining epithelium in human small intestine.
Panel C : anti-MUC2 staining goblet cells in human small intestine.
Panel D : anti-REG1B staining Paneth cells in human small intestine.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab316971, ab272692 and ab323315 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-REG1B antibody [EPR28864-4] (AB323315)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling REG1B with ab323315 at 1/5000 (0.106 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human skeletal muscle (PMID : 8110835).
The section was incubated with ab323315 for 10 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-REG1B antibody [EPR28864-4] (AB323315)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling REG1B with ab323315 at 1/5000 (0.106 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human spleen.
The section was incubated with ab323315 for 10 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-REG1B antibody [EPR28864-4] (AB323315)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human ileum tissue staining BTNL3 with ab324982 at a 1/500 (0.106 μg/ml) dilution, ab272692 anti-MUC2 used at a 1/2000 (0.26 μg/ml) dilution and ab323315 anti-REG1B used at a 1/5000 (0.972 μg/ml) dilution.

Panel A : anti-BTNL3 (green; Opal™570), anti-MUC2 (magenta; Opal™690), anti-REG1B (gray; Opal™520) on human ileum.

Panel B : anti-BTNL3 staining brush border in human ileum.

Panel C : anti-MUC2 staining goblet cells in human ileum.

Panel D : anti-REG1B staining Paneth cells in human ileum.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324982, ab272692 and ab323315 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 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-REG1B antibody [EPR28864-4] (AB323315)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling REG1B with ab323315 at 1/5000 (0.106 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human pancreas (PMID : 8110835 ).
The section was incubated with ab323315 for 10 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• WB

Supplier Data

Western blot - Anti-REG1B antibody [EPR28864-4] (AB323315)

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

Negative control : spleen, breast.

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

All lanes:

Western blot - Anti-REG1B antibody [EPR28864-4] (ab323315) at 1/1000 dilution

Lane 1:

Human pancreas tissue lysate at 20 µg

Lane 2:

Human spleen tissue lysate at 20 µg

Lane 3:

Human breast 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: 19 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-REG1B antibody [EPR28864-4] (AB323315)

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

This antibody does not cross-react with human REG1A.

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

All lanes:

Western blot - Anti-REG1B antibody [EPR28864-4] (ab323315) at 1/1000 dilution

Lane 1:

His-tagged human REG1B recombinant protein at 10 ng

Lane 2:

His-tagged human REG1A recombinant protein 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: 19 kDa

false

Exposure time: 3s