Anti-PLOD2/LH2 antibody [EPR25160-20]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PLOD2/LH2 antibody [EPR25160-20] (AB313765)

Immunohistochemical analysis of paraffin-embedded (A) Wild-type HEK293T tissue labeling PLOD2/LH2 with ab313765 at 1/2000 (0.252 µg/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 HEK293T cell pellet. The section was incubated with ab313765 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PLOD2/LH2 antibody [EPR25160-20] (AB313765)

Immunohistochemical analysis of paraffin-embedded Human hepatocellular tissue labeling PLOD2/LH2 with ab313765 at 1/2000 (0.252 µg/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 ab313765 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 [EPR25160-20] (AB313765)

Immunohistochemical analysis of paraffin-embedded Human clear cell renal tissue labeling PLOD2/LH2 with ab313765 at 1/2000 (0.252 µg/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 ab313765 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

• IP

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Immunoprecipitation - Anti-PLOD2/LH2 antibody [EPR25160-20] (AB313765)

PLOD2/LH2 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab313765 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313765 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 : ab313765 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313765 in HeLa whole cell lysate. Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 15 seconds

All lanes:

Immunoprecipitation - Anti-PLOD2/LH2 antibody [EPR25160-20] (ab313765) 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='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>)

false

Exposure time: 15s

• IP

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Immunoprecipitation - Anti-PLOD2/LH2 antibody [EPR25160-20] (AB313765)

PLOD2/LH2 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab313765 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313765 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate Lane 2 : ab313765 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313765 in NIH/3T3 whole cell lysate. Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 180 seconds

All lanes:

Immunoprecipitation - Anti-PLOD2/LH2 antibody [EPR25160-20] (ab313765) at 1/30 dilution

All lanes:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Secondary

All lanes:

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

false

Exposure time: 180s

• WB

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Western blot - Anti-PLOD2/LH2 antibody [EPR25160-20] (AB313765)

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

Lysates at 20 µg per lane.

The samples were run on a Bis-Tris gel under reducing conditions. Western blot : Anti-PLOD2/LH2 antibody (ab313765) 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, ab313765 was shown to bind specifically to PLOD2/LH2.

Target of interest was observed at 95 kDa in wild-type Hela 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 [EPR25160-20] (ab313765) at 1/1000 dilution

Lane 1:

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

Lane 2:

Western blot - Human PLOD2 (PLOD2/LH2) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-plod2-plod2-lh2-knockout-hek-293t-cell-line-ab266279'>ab266279</a>)

Lane 2:

Western blot - Human PLOD2 (PLOD2/LH2) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-plod2-plod2-lh2-knockout-hek-293t-cell-lysate-ab258591'>ab258591</a>) 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

• WB

Supplier Data

Western blot - Anti-PLOD2/LH2 antibody [EPR25160-20] (AB313765)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : 81 seconds

All lanes:

Western blot - Anti-PLOD2/LH2 antibody [EPR25160-20] (ab313765) 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 [EPR25160-20] (AB313765)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : Lane 1-3 : 158 seconds, Lane 4-5 : 48 seconds

All lanes:

Western blot - Anti-PLOD2/LH2 antibody [EPR25160-20] (ab313765) at 1/1000 dilution

Lane 1:

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

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Lane 4:

Mouse testis tissue lysate at 20 µg

Lane 5:

C2C12 (mouse myoblast) 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 1/20000 dilution

Observed band size: 95 kDa

false

Exposure time: 158s

• WB

Supplier Data

Western blot - Anti-PLOD2/LH2 antibody [EPR25160-20] (AB313765)

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 : 26 seconds

All lanes:

Western blot - Anti-PLOD2/LH2 antibody [EPR25160-20] (ab313765) 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

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

Observed band size: 95 kDa

false

Exposure time: 26s

• Dot

Supplier Data

Dot Blot - Anti-PLOD2/LH2 antibody [EPR25160-20] (AB313765)

Dot blot analysis of PLOD2/LH2 using ab313765 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