Anti-PLOD2/LH2 antibody [EPR25160-22] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PLOD2/LH2 antibody [EPR25160-22] - BSA and Azide free (AB314641)

This data was developed using ab314640, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded (A) Wild-type HEK293T (human embryonic kidney epithelial cell) cell pellet (B) PLOD2 knockout HEK293T cell pellet tissue labeling PLOD2/LH2 with ab314640 at 1/2000 (0.257 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on (A) Wild-type HEK293T cell pellet, no staining on (B) PLOD2 knockout KEK293T cell pellet.The section was incubated with ab314640 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-PLOD2/LH2 antibody [EPR25160-22] - BSA and Azide free (AB314641)

This data was developed using ab314640, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma and adjacent tissue tissue labeling PLOD2/LH2 with ab314640 at 1/2000 (0.257 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on (A) human hepatocellular carcinoma tissue and (B) very weak staining on the adjacent tissue (PMID : 33014843). The section was incubated with ab314640 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-PLOD2/LH2 antibody [EPR25160-22] - BSA and Azide free (AB314641)

This data was developed using ab314640, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human clear cell renal cell carcinoma and adjacent tissue tissue labeling PLOD2/LH2 with ab314640 at 1/2000 (0.257 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on (A) human clear cell renal cell carcinoma tissue and (B) very weak staining on the adjacent tissue (PMID : 33287763). The section was incubated with ab314640 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-PLOD2/LH2 antibody [EPR25160-22] - BSA and Azide free (AB314641)

This data was developed using ab314640, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling PLOD2/LH2 with ab314640 at 1/50 (10.28 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1010 (2ug/ml) dilution (Green). Confocal image showing cytoplasmic staining colocalized with endoplasmic reticulum marker in Hela cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Anti-KDEL mouse monoclonal antibody was used to counterstain at 1/100 (1 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1010 (2ug/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PLOD2/LH2 antibody [EPR25160-22] - BSA and Azide free (AB314641)

This data was developed using ab314640, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PLOD2 KO HEK293T (human PLOD2 knockout embryonic kidney epithelial cell, Left) / Parental HEK293T (Right) cells labelling PLOD2/LH2 with ab314640 at 1/20000 dilution (0.0025 ug, Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-PLOD2/LH2 antibody [EPR25160-22] - BSA and Azide free (AB314641)

This data was developed using ab314640, the same antibody clone in a different buffer formulation. PLOD2/LH2 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab314640 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314640 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate Lane 2 : ab314640 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314640 in HeLa whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-PLOD2/LH2 antibody [EPR25160-22] (<a href='/ko/products/primary-antibodies/plod2-lh2-antibody-epr25160-22-ab314640'>ab314640</a>) at 1/30 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/ko/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-PLOD2/LH2 antibody [EPR25160-22] - BSA and Azide free (AB314641)

This data was developed using ab314640, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The identity of the band around 37 kDa is unknown. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PLOD2/LH2 antibody [EPR25160-22] (<a href='/ko/products/primary-antibodies/plod2-lh2-antibody-epr25160-22-ab314640'>ab314640</a>) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human colon cancer tissue lysate at 20 µg

Lane 3:

Human liver cancer 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: 95 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-PLOD2/LH2 antibody [EPR25160-22] - BSA and Azide free (AB314641)

This data was developed using ab314640, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. Exposure time : Lane1-3 : 70 seconds; Lane 4 : 180 seconds

All lanes:

Western blot - Anti-PLOD2/LH2 antibody [EPR25160-22] (<a href='/ko/products/primary-antibodies/plod2-lh2-antibody-epr25160-22-ab314640'>ab314640</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

Hela transfected with siRNA specifically targeting PLOD2 whole cell lysate at 20 µg

Lane 3:

DU 145 (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 95 kDa

false

• WB

Supplier Data

Western blot - Anti-PLOD2/LH2 antibody [EPR25160-22] - BSA and Azide free (AB314641)

This data was developed using ab314640, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates at 20 µg per lane. Western blot : Anti-PLOD2/LH2 antibody (ab314640) staining at 1/1000 dilution, shown in green;Mouse anti-GAPDH antibody 6C5 (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab314640 was shown to bind specifically to PLOD2/LH2. Target of interest was observed at 95 kDa in wild-type HEK293T cell lysates (lane 1) with no signal observed at this size in PLOD2 knockout cell line ab266279 (knockout cell lysate ab258591) (lane 2). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.

All lanes:

Western blot - Anti-PLOD2/LH2 antibody [EPR25160-22] (<a href='/ko/products/primary-antibodies/plod2-lh2-antibody-epr25160-22-ab314640'>ab314640</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

PLOD2 knockout HEK293T whole cell lysate at 20 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (800CW) and Goat Anti-Mouse IgG H&L (680RD) at 1/20000 dilution

Observed band size: 95 kDa

false

• Dot

Supplier Data

Dot Blot - Anti-PLOD2/LH2 antibody [EPR25160-22] - BSA and Azide free (AB314641)

This data was developed using ab314640, the same antibody clone in a different buffer formulation. Dot blot analysis of PLOD2/LH2 using ab314640 at 1 : 1000 (0.503 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution. Exposure time : 180 seconds Blocking and diluting buffer and concentration : 5% NFDM/TBST