Anti-GAPDH antibody [EPR16891] - Loading Control

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocyte of rat spleen is observed. Counter stained with Hematoxylin.

Negative control : Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Immunocytochemistry/immunofluorescence staining of 4% paraformaldehyde fixed; 0.1% triton X 100 permeabilized HeLa (human cervix adenocarcinoma) cells labeling GAPDH with ab181602 at dilution of 1/500. The secondary antibody used was Alexa Fluor^® 488; goat anti-rabbit IgG (ab150077) at a dilution of 1/400. Nucleus was counter-stained with DAPI (blue). ab7291, a mouse anti-tubulin antibody (1/500) was used to stain tubulin along with ab150120 (AlexaFluor^®594 goat anti-mouse secondary, 1/500). The negative controls are shown in the bottom middle and right hand panels- for negative control 1 primary antibody (ab181602; 1/500) and secondary antibody (ab150120; 1/500) was used. For negative control 2 primary antibody (ab7291; 1/500) and secondary antibody (ab150077; 1/400) was used.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Immunohistochemical analysis of paraffin-embedded human transitional cell carcinoma of bladder tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic and nucleus staining on the tumor cells of transitional cell carcinoma of human bladder is observed. Counter stained with Hematoxylin.

Negative control : Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling GAPDH with ab181602 at 1/180 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• IP

Lab

Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

STT3A was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney epithelial cell) whole cell lysate with ab320831 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab320831 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

In Immunoprecipitation, ab320831 was shown to bind specifically to STT3A. Target of interest was enriched at 60 kDa in wild-type HEK-293T cell lysates (lane 2) with no signal observed at this size in STT3A knockout cell line (lane 5) (lane 5, knockout cell line ab266320 / knockout cell lysate ab259164).
The bands above 60kDa are likely to be aggregation, and are absent in the KO samples. Unlike the Western Blot data, these samples have been boiled. ab181602 was used as a GAPDH loading control.

All lanes:

Immunoprecipitation - Anti-STT3A antibody [EPR29178-10] (<a href='/en-us/products/primary-antibodies/stt3a-antibody-epr29178-10-ab320831'>ab320831</a>) at 1/1000 dilution

Lane 1:

Parental HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 10 µg

Lane 2:

Parental HEK-293T (human embryonic kidney epithelial cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/stt3a-antibody-epr29178-10-ab320831'>ab320831</a> in Parental HEK-293T whole cell lysate

Lane 4:

STT3A KO HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 10 µg

Lane 5:

Immunoprecipitation - Human STT3A knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-stt3a-knockout-hek-293t-cell-line-ab266320'>ab266320</a>)

Lane 6:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/stt3a-antibody-epr29178-10-ab320831'>ab320831</a> in STT3A KO HEK-293T 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

Observed band size: 60 kDa

false

Exposure time: 84s

• IP

Supplier Data

Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Dnmt3a was immunoprecipitated from 0.35 mg Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab323708 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323708 at 1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Dnmt3a antibody [EPR29184-89] (<a href='/en-us/products/primary-antibodies/dnmt3a-antibody-epr29184-89-ab323708'>ab323708</a>) at 1/1000 dilution

Lane 1:

(Input) Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/dnmt3a-antibody-epr29184-89-ab323708'>ab323708</a> at 1/30 IP in Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/dnmt3a-antibody-epr29184-89-ab323708'>ab323708</a> in parental HeLa whole cell lysate at 10 µg

Lane 4:

(Input) DNMT3A knockout HeLa whole cell lysate at 10 µg

Lane 5:

<a href='/en-us/products/primary-antibodies/dnmt3a-antibody-epr29184-89-ab323708'>ab323708</a> at 1/30 IP in DNMT3A knockout HeLa whole cell lysate at 10 µg

Lane 6:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/dnmt3a-antibody-epr29184-89-ab323708'>ab323708</a> in DNMT3A knockout HeLa whole cell lysate at 10 µg

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

Observed band size: 130 kDa,36 kDa

false

Exposure time: 24s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocytes of mouse spleen is observed. Counter stained with Hematoxylin.

Negative control : Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : SW480 (PMID : 30266650).

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

All lanes:

Western blot - Anti-SMP30 antibody [EPR29674-517] (<a href='/en-us/products/primary-antibodies/smp30-antibody-epr29674-517-ab324949'>ab324949</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

SW480 (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='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 35 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : A375,

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

All lanes:

Western blot - Anti-ANGPTL3 antibody [EPR30236-518] (<a href='/en-us/products/primary-antibodies/angptl3-antibody-epr30236-518-ab324272'>ab324272</a>) at 1/1000 dilution

Lane 1:

Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 100 µg

Lane 2:

A375 (human malignant melanoma epithelial cell) whole cell lysate at 100 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/1000 dilution

Observed band size: 53 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : Jurkat (PMID : 11980907), T-47D, JAR.

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

Exposure time : Lanes 1-6 : 70 seconds; Lane 7 : 180 seconds.

All lanes:

Western blot - Anti-AMID antibody [EPR29828-39] (<a href='/en-us/products/primary-antibodies/amid-antibody-epr29828-39-ab324520'>ab324520</a>) at 1/1000 dilution

Lane 1:

DLD-1 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

Lane 3:

786-O (human kidney epithelial cell) whole cell lysate at 20 µg

Lane 4:

Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 5:

JAR (human placenta choriocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

A-172 (human brain glioblastoma cell ) whole cell lysate at 20 µg

Lane 7:

HepG2 (human hepatocellar 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: 41 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : MCF7 and SK-BR-3.

In Western blot, Anti-GAPDH antibody [EPR16891] (ab181602) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Glycerol 3 Phosphate Dehydrogenase antibody [EPR25996-4] (<a href='/en-us/products/primary-antibodies/glycerol-3-phosphate-dehydrogenase-antibody-epr25996-4-ab313871'>ab313871</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Human skeletal muscle cytoplasma tissue lysate at 20 µg

Lane 5:

human liver tissue lysate at 20 µg

Lane 6:

Mouse liver tissue lysate at 20 µg

Lane 7:

Rat liver 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 1/20000 dilution

Observed band size: 35 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

In Western blot, Anti-COX IV antibody [EPR9442 (ABC)] - Mitochondrial Loading Control - (ab202554) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-PDHA1 (phospho S232) antibody [EPR29825-102] (<a href='/en-us/products/primary-antibodies/pdha1-phospho-s232-antibody-epr29825-102-ab325169'>ab325169</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) mitochondria fraction at 20 µg

Lane 2:

HeLa non-mitochondrial fraction 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: 43 kDa,17 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 32596239; PMID : 23746844).

The identity of the lower MW band at approximately 30 kDa is unknown.

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

All lanes:

Western blot - Anti-Cdk8 antibody [EPR26566-270] (<a href='/en-us/products/primary-antibodies/cdk8-antibody-epr26566-270-ab325133'>ab325133</a>) at 1/1000 dilution

Lane 1:

Human colon tissue lysate at 48 µg

Lane 2:

Human colon cancer tissue lysate at 48 µg

Lane 3:

Human pancreas tissue lysate at 48 µg

Lane 4:

Human pancreas cancer tissue lysate at 48 µg

Lane 5:

Human testis tissue lysate at 48 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 53 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : human lung (PMID : 10644446).

The band around 53 kDa is the full length form ANGPTL3 and 30 kDa the cleaved C-terminal fibrinogen-like domain (ANGPTL3-FLD).

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

All lanes:

Western blot - Anti-ANGPTL3 antibody [EPR30236-518] (<a href='/en-us/products/primary-antibodies/angptl3-antibody-epr30236-518-ab324272'>ab324272</a>) at 1/1000 dilution

Lane 1:

Human liver cancer tissue lysate at 100 µg

Lane 2:

Human liver tissue lysate at 100 µg

Lane 3:

Human lung tissue lysate at 100 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/1000 dilution

Observed band size: 53 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000000 dilution.

In Western blot, Anti-Myc tag antibody [9E10] - Loading Control (ab32) staining at 1/10000 dilution.

In Western blot, Anti-6X His tag® antibody [EPR20547] - Loading Control (ab213204) staining at 1/10000 dilution.

This antibody does not cross react with DR5 and DcR2.

All lanes:

Western blot - Anti-DR4 antibody [EPR28152-57] (<a href='/en-us/products/primary-antibodies/dr4-antibody-epr28152-57-ab312848'>ab312848</a>) at 1/1000 dilution

Lane 1:

293T (Human embryonic kidney epithelial cell) transfected with an empty vector (vector control), containing a myc-His- tag® whole cell lysate at 20 µg

Lane 2:

293T transfected with human DR4 expression vector containing a myc- His-tag®, whole cell lysate at 20 µg

Lane 3:

293T transfected with human DcR2 expression vector containing a myc- His-tag®, whole cell lysate at 48 µg

Lane 4:

Recombinant human DR5 fragment protein containing a His-tag®, whole cell lysate 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: 48 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression or negative samples : MOLT-4, Jurkat (PMID : 24141767, PMID : 26121639).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000000 dilution.

All lanes:

Western blot - Anti-Human IgM antibody [EPR5539-65-4] (<a href='/en-us/products/primary-antibodies/human-igm-antibody-epr5539-65-4-ab134159'>ab134159</a>) at 1/1000 dilution

Lane 1:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 2:

Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 3:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Lane 4:

Jurkat (human T cell leukemia T lymphocyte) 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: 75 kDa

false

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Performed under reducing conditions.

In Western blot, ab313339 was shown to bind specifically to CD73. A band was observed at 68 kDa in wild-type A431 cell lysates whereas loss of signal was observed in the CD73 knockout cell line.

In Western blot, anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/10000 dilution.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-CD73 antibody [EPR28213-52] (<a href='/en-us/products/primary-antibodies/cd73-antibody-epr28213-52-ab313339'>ab313339</a>) at 1/1000 dilution

Lane 1:

Wild-type A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 2:

CD73 knockout A431 whole cell 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: 63 kDa

Observed band size: 68 kDa

true

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : spleen, lung (PMID : 11481335).

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

Exposure time : Lanes 1-3 : 48 seconds; Lane 4 : 180 seconds.

All lanes:

Western blot - Anti-DGAT2 antibody [EPR27385-45] (<a href='/en-us/products/primary-antibodies/dgat2-antibody-epr27385-45-ab325121'>ab325121</a>) at 1/1000 dilution

Lane 1:

Human liver tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Lane 3:

Human spleen tissue lysate at 20 µg

Lane 4:

Human fat tissue lysate at 20 µg

Secondary

All lanes:

Western blot at 1/2000 dilution

Observed band size: 37 kDa

false

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : IMR-90 (PMID : 24742710).

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

All lanes:

Western blot - Anti-Rad51C antibody [EPR30494-537] (<a href='/en-us/products/primary-antibodies/rad51c-antibody-epr30494-537-ab325261'>ab325261</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 2:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 50 µg

Lane 3:

IMR-90 (human lung fibroblast) whole cell lysate at 50 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: 38 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Negative control : A549, HepG2.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11739566; PMID : 22112368; PMID : 14997013).

The identity of the higher MW band at approximately 50 kDa (in lane 2) is unknown.

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

All lanes:

Western blot - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (<a href='/en-us/products/primary-antibodies/alpha-synuclein-n-term-antibody-epr29522-6-ab324977'>ab324977</a>) at 1/1000 dilution

Lane 1:

G-401 (human rhabdoid tumor kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

SK-MEL-28 (human malignant melanoma cell ) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot at 1/20000 dilution

Observed band size: 18 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

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

All lanes:

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (<a href='/en-us/products/primary-antibodies/adenine-nucleotide-translocator-2-ant-2-antibody-epr29813-534-ab325120'>ab325120</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 SLC25A5 whole cell lysate at 20 µg

Lane 3:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 6:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

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

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 32 kDa,36 kDa

false

Exposure time: 125s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Negative control : SH-SY5Y (PMID : 30650643 and PMID : 27345400). Spleen (PMID : 24309898).

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

Exposure time : Lanes 1-2 : 4 seconds; Lanes 3-4 : 37 seconds

All lanes:

Western blot - Anti-Cytokeratin 19 antibody [EP1580Y] - Rat IgG2a (Chimeric) (<a href='/en-us/products/primary-antibodies/cytokeratin-19-antibody-ep1580y-rat-igg2a-chimeric-ab323561'>ab323561</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 40 µg

Lane 3:

Human colon tissue lysate at 20 µg

Lane 4:

Human spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG Fc (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-fc-hrp-preadsorbed-ab97090'>ab97090</a>) at 1/20000 dilution

Observed band size: 44 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Performed under reducing conditions.

In Western blot, ab325169 was shown to bind specifically to PDHA1. Target of interest was observed at 43 kDa in induced wild-type A549 cell lysates (lane 2) with no signal observed at this size in induced PDHA1 knockout cell line (lane 4).

Lanes 1-4 on the left untreated membrane with Lambda Protein Phosphatase; Lanes 1-4 on the right treated membrane with Lambda Protein Phosphatase.

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

In Western blot, Anti-PDHA1 antibody [EPR11098] - (ab168379) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-PDHA1 (phospho S232) antibody [EPR29825-102] (<a href='/en-us/products/primary-antibodies/pdha1-phospho-s232-antibody-epr29825-102-ab325169'>ab325169</a>) at 1/1000 dilution

Lane 1:

Untreated A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

A549 treated with 100ng/ml Calyculin A for 30 minutes whole cell lysate at 20 µg

Lane 3:

Untreated PDHA1 knockout A549 whole cell lysate at 20 µg

Lane 4:

PDHA1 knockout A549 treated with 100ng/ml Calyculin A for 30 minutes 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: 43 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

In Western blot, ab324977 was shown to bind specifically to Alpha-synuclein. Target of interest was observed at 18 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in Alpha-synuclein knockout cell line (lane 2).

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

All lanes:

Western blot - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (<a href='/en-us/products/primary-antibodies/alpha-synuclein-n-term-antibody-epr29522-6-ab324977'>ab324977</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate at 20 µg

Lane 2:

Alpha-synuclein knockout HAP1 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot at 1/20000 dilution

Observed band size: 18 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000000 dilution.

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

Low expression or negative samples : skeletal muscle, MCF7, T-47D (PMID : 10945981, PMID : 35082382).

All lanes:

Western blot - Anti-SEP15 antibody [NCIR128A] (<a href='/en-us/products/primary-antibodies/sep15-antibody-ncir128a-ab124840'>ab124840</a>) at 1/1000 dilution

Lane 1:

Mouse lung tissue lysate at 20 µg

Lane 2:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 3:

Rat skeletal muscle tissue lysate at 20 µg

Lane 4:

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

Lane 5:

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

Lane 6:

T-47D (human ductal breast epithelial tumor 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: 15 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

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

Low expression : liver, testis and spleen.

All lanes:

Western blot - Anti-ATP1B2 antibody [EPR15461(B)] - C-terminal (<a href='/en-us/products/primary-antibodies/atp1b2-antibody-epr15461b-c-terminal-ab185207'>ab185207</a>) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human testis tissue lysate at 20 µg

Lane 4:

Human spleen 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 1/20000 dilution

Observed band size: 40-50 kDa,36 kDa

false

Exposure time: 1s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Performed under reducing conditions.

In Western blot, ab325133 was shown to bind specifically to Cdk8. Target of interest was observed at 53 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in Cdk8 knockout cell line (lane 2).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 32596239; PMID : 23746844).

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

All lanes:

Western blot - Anti-Cdk8 antibody [EPR26566-270] (<a href='/en-us/products/primary-antibodies/cdk8-antibody-epr26566-270-ab325133'>ab325133</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate at 20 µg

Lane 2:

CDK8 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 6:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 7:

HeLa (human cervical adenocarcinoma 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: 53 kDa,36 kDa

false

Exposure time: 81s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Lanes 1 - 4:

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (<a href='/en-us/products/primary-antibodies/trka-phospho-y674trkb-phospho-y706trkc-phospho-y709-antibody-epr22298-67-ab229908'>ab229908</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-TrkA (pY674) + TrkB (pY706) + TrkC (pY709) antibody [EPR22298-67] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/trka-phospho-y674trkb-phospho-y706trkc-phospho-y709-antibody-epr22298-67-bsa-and-azide-free-ab264070'>ab264070</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human TrkA expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human TrkB expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human TrkC expression vector containing a His-tag, 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: 140 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : A549, HT-1080(PMID : 25006253), colon

The expression profile observed is consistent with what has been described in the literature(PMID : 25006253)

This antibody can recognize both the full-length PTK7 and the N-terminal PTK7. The 160 kDa form may represent the full-length protein with glycosylation modifications, while the 110 kDa form likely corresponds to the N-terminal PTK7 glycosylation modification.

The identity of the bands lower than 37kDa are unknown.

Lanes 1-6 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 and lanes 7-9 are incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

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

All lanes:

Western blot - Anti-CCK4 antibody [EPR30150-507] (<a href='/en-us/products/primary-antibodies/cck4-antibody-epr30150-507-ab325170'>ab325170</a>) at 1/1000 dilution

Lane 1:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HT-29 (human colorectal adenocarcinoma epithelial cell) 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

Lane 5:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

Human lung tissue lysate at 20 µg

Lane 8:

Human colon tissue lysate at 20 µg

Secondary

Lanes 1 - 6:

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

Lanes 7 - 8:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 110 kDa,160 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 28541286 and PMID : 157438355).

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

Exposure time : Lanes 1-2 : 147 seconds; Lanes 3-4 : 26 seconds

All lanes:

Western blot - Anti-PSAP antibody [RM1353] (<a href='/en-us/products/primary-antibodies/psap-antibody-rm1353-ab324885'>ab324885</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 PSAP whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 4:

NIH/3T3 transfected with siRNA specifically targeting PSAP 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 1/200000 dilution

Observed band size: 60-80 kDa,12 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : liver

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

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR29833-94] (<a href='/en-us/products/primary-antibodies/calpain-2-antibody-epr29833-94-ab325140'>ab325140</a>) at 1/1000 dilution

Lane 1:

Human stomach tissue lysate at 20 µg

Lane 2:

Human tonsil tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Lane 4:

Human liver cancer tissue lysate at 20 µg

Secondary

All lanes:

Western blot at 1/2000 dilution

Observed band size: 76 kDa,36 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : MCF7.

To minimize protein degradation, cells/tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

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

All lanes:

Western blot - Anti-SALL1 antibody [EPR28911-52] (<a href='/en-us/products/primary-antibodies/sall1-antibody-epr28911-52-ab325098'>ab325098</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Secondary

All lanes:

Western blot at 1/20000 dilution

Observed band size: 200 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

The β chain undergoes degradation, resulting in lower molecular weight fragments (PMID : 32522525). To minimize protein degradation, cells/tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting right away.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000000 dilution.

All lanes:

Western blot - Anti-Met (c-Met) antibody [SP44] - C-terminal (<a href='/en-us/products/primary-antibodies/met-c-met-antibody-sp44-c-terminal-ab227637'>ab227637</a>) at 1/1000 dilution

Lane 1:

Fresh HeLa cell lysate at 20 µg

Lane 2:

Frozen HeLa 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: 145-175 kDa,35-55 kDa

false

Exposure time: 60s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

c-Met can undergo glycosylation, resulting in a higher molecular weight. It can also be cleaved into α and β chains (PMID : 34210960). The β chain undergoes degradation, resulting in lower molecular weight fragments (PMID : 32522525). This antibody detects both the full-length c-Met and the β chains.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000000 dilution.

All lanes:

Western blot - Anti-Met (c-Met) antibody [SP44] - C-terminal (<a href='/en-us/products/primary-antibodies/met-c-met-antibody-sp44-c-terminal-ab227637'>ab227637</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MET knockout HeLa 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: 145-175 kDa,35-55 kDa

false

Exposure time: 60s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

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

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR29833-94] (<a href='/en-us/products/primary-antibodies/calpain-2-antibody-epr29833-94-ab325140'>ab325140</a>) at 1/1000 dilution

Lane 1:

PC-3 (human prostate adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 1 µg

Lane 2:

PC-3 transfected with siRNA specifically targeting CAPN2 whole cell lysate at 1 µg

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 76 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : LNCaP, Jurkat

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

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR29833-94] (<a href='/en-us/products/primary-antibodies/calpain-2-antibody-epr29833-94-ab325140'>ab325140</a>) at 1/1000 dilution

Lane 1:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 76 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : Ramos, small intestine (PMID : 155411360), lung, cerebellum.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 23870553).

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

All lanes:

Western blot - Anti-Metallothionein antibody [EPR29104-22] (<a href='/en-us/products/primary-antibodies/metallothionein-antibody-epr29104-22-ab325177'>ab325177</a>) at 1/1000 dilution

Lane 1:

MG-63 (human osteosarcoma fibroblast) transfected with scrambled siRNA control whole cell lysate at 50 µg

Lane 2:

MG-63 transfected with siRNA specifically targeting Metallothionein whole cell lysate at 50 µg

Lane 3:

Ramos (human Burkitt's lymphoma B lymphocyte) at 50 µg

Lane 4:

Human liver tissue lysate at 50 µg

Lane 5:

Human small intestine tissue lysate at 50 µg

Lane 6:

Human lung tissue lysate at 50 µg

Lane 7:

Human cerebellum tissue lysate at 50 µg

Secondary

All lanes:

Western blot at 1/2000 dilution

Observed band size: 15-20 kDa,36 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : liver (PMID : 24742710).

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

The identity of the bands above 50 kDa and below 25kda (in lane 1) are unknown.

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

All lanes:

Western blot - Anti-Rad51C antibody [EPR30494-537] (<a href='/en-us/products/primary-antibodies/rad51c-antibody-epr30494-537-ab325261'>ab325261</a>) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 50 µg

Lane 2:

Human liver tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 38 kDa,36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

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

All lanes:

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (<a href='/en-us/products/primary-antibodies/adenine-nucleotide-translocator-2-ant-2-antibody-epr29813-534-ab325120'>ab325120</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa non-mitochondrial fraction at 20 µg

Lane 3:

HeLa mitochondrial fraction at 20 µg

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 32 kDa,36 kDa,16 kDa

false

Exposure time: 125s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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 : Lanes 1-3 : 180 seconds; Lanes 4-5 : 70 seconds.

All lanes:

Western blot - Anti-DGAT2 antibody [EPR27385-45] (<a href='/en-us/products/primary-antibodies/dgat2-antibody-epr27385-45-ab325121'>ab325121</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HepG2 transfected with siRNA specifically targeting DGAT2 whole cell lysate at 20 µg

Lane 3:

PAZ6 (human adipose epithelial cell) whole cell lysate at 20 µg

Lane 4:

Untreated 3T3-L1 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

3T3-L1 differentiated for 6 days whole cell lysate at 20 µg

Secondary

All lanes:

Western blot at 1/20000 dilution

Observed band size: 37 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

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

Low expression : liver, kidney(PMID : 8194594)

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11739566; PMID : 22112368; PMID : 14997013).

Lanes 1-4 are incubated with are incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 and lanes 5-8 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000.

The identity of the higher MW band at approximately 50 kDa (in lane 2, 5 and lanes 7- 8) is unknown.

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

All lanes:

Western blot - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (<a href='/en-us/products/primary-antibodies/alpha-synuclein-n-term-antibody-epr29522-6-ab324977'>ab324977</a>) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human hypothalamus tissue lysate at 20 µg

Lane 4:

Human thalamus tissue lysate at 20 µg

Lane 5:

Mouse brain tissue lysate at 20 µg

Lane 6:

Mouse liver tissue lysate at 20 µg

Lane 7:

Mouse hippocampus tissue lysate at 20 µg

Lane 8:

Mouse kidney tissue lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Lanes 5 - 8:

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: 18 kDa,36 kDa

false

Exposure time: 103s