Anti-OLFM4 antibody [EPR29030-1]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-OLFM4 antibody [EPR29030-1] (AB322350)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling OLFM4 with ab322350 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human testis (PMID : 25364714).
The section was incubated with ab322350 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-OLFM4 antibody [EPR29030-1] (AB322350)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling OLFM4 with ab322350 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human skeletal muscle (PMID : 25364714).
The section was incubated with ab322350 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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-OLFM4 antibody [EPR29030-1] (AB322350)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Capan-1 (human pancreatic adenocarcinoma epithelial cell) cells labelling OLFM4 with ab322350 at 1/100 (5.04 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in Capan-1 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : MCF7
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Magenta).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-OLFM4 antibody [EPR29030-1] (AB322350)

Immunohistochemical analysis of paraffin-embedded Human appendix tissue labeling OLFM4 with ab322350 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human appendix.
The section was incubated with ab322350 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-OLFM4 antibody [EPR29030-1] (AB322350)

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling OLFM4 with ab322350 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon cancer.
The section was incubated with ab322350 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-OLFM4 antibody [EPR29030-1] (AB322350)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling OLFM4 with ab322350 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon (PMID : 19450592).
The section was incubated with ab322350 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-OLFM4 antibody [EPR29030-1] (AB322350)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling OLFM4 with ab322350 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen.
The section was incubated with ab322350 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-OLFM4 antibody [EPR29030-1] (AB322350)

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

Negative control : MCF7, U-937

The identity of MW band at approximately 35kDa (in lane 1) is unknown.

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

All lanes:

Western blot - Anti-OLFM4 antibody [EPR29030-1] (ab322350) at 1/1000 dilution

Lane 1:

Capan-1 (human pancreatic adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

U-937 (human histiocytic lymphoma monocyte) 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 1/100000 dilution

Observed band size: 70-78 kDa,36 kDa

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-OLFM4 antibody [EPR29030-1] (AB322350)

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

Low expression : testis, pancreas, heart (PMID : 25364714).

The identity of the bands lower than 50 kDa (in lane1, 4 and 5) are unknown.

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

Exposure time : Lanes 1-3 : 6 seconds; Lanes 4-5 : 48 seconds

All lanes:

Western blot - Anti-OLFM4 antibody [EPR29030-1] (ab322350) at 1/1000 dilution

Lane 1:

Human small intestine tissue lysate at 20 µg

Lane 2:

Human testis tissue lysate at 20 µg

Lane 3:

Human pancreas tissue lysate at 20 µg

Lane 4:

Human colon tissue lysate at 20 µg

Lane 5:

Human heart 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: 70-78 kDa,36 kDa

false