Goat Anti-Rabbit IgG H&L (HRP)

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Blocking buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-RAB29 (phospho T71) antibody [MJF-R24-17-1] (<a href='/ko/products/primary-antibodies/rab29-phospho-t71-antibody-mjf-r24-17-1-ab241062'>ab241062</a>) at 1/5000 dilution

Lane 1:

HEK-293 (human embryonic kidney) over-expressing HA-tagged RAB29 and LRRK2 [Y1699C], whole cell lysate at 15 µg

Lane 2:

MLi-2 (100nM for 60min) treated HEK-293 over-expressing HA-tagged RAB29 and LRRK2 [Y1699C], whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 23 kDa

Observed band size: 23 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Immunohistochemical analysis of paraffin-embedded Human skin tissue labeling Histone H3 (mono methyl K79) with ab177183 at 1/5000 dilution, followed by ab97051 Goat Anti-Rabbit HRP (IgG H&L) at 1/500 dilution. Nucleus staining on Human skin tissue 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Immunohistochemical staining of paraffin embedded human endometrial carcinoma with purified ab32063 at a working dilution of 1 in 200. The secondary antibody used is ab97051 a HRP goat anti-rabbit IgG (H+L) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer pH 9.0.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

ab92511 staining HLA-DR in human spleen tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/700. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500. Negative control 1 : PBS in place of primary antibody.

• IP

Supplier Data

Immunoprecipitation - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

DDR1 (phospho Y796) + DDR2 (phospho Y740) was immunoprecipitated from 0.35 mg T-47D treated with 20 ug/ml Collagen I for 18 hours whole cell lysate with ab322921 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322921 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-DDR1 (phospho Y796) + DDR2 (phospho Y740) antibody [EPR28699-33] (<a href='/ko/products/primary-antibodies/ddr1-phospho-y796-ddr2-phospho-y740-antibody-epr28699-33-ab322921'>ab322921</a>) at 1/1000 dilution

Lane 1:

T-47D treated with 20 ug/ml Collagen I for 18 hours whole cell lysate at 10 µg

Lane 2:

<a href='/ko/products/primary-antibodies/ddr1-phospho-y796-ddr2-phospho-y740-antibody-epr28699-33-ab322921'>ab322921</a> at 1/30 IP in T-47D treated with 20 ug/ml Collagen I for 18 hours whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/ko/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/ko/products/primary-antibodies/ddr1-phospho-y796-ddr2-phospho-y740-antibody-epr28699-33-ab322921'>ab322921</a> in T-47D treated with 20 ug/ml Collagen I for 18 hours whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution

Observed band size: 125 kDa

false

Exposure time: 76s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

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

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and IL1r1-KO homozygous mice (Strain ID : T005977).

All lanes:

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL1 Receptor I/IL-1R-1 antibody [EPR22198-36] (<a href='/ko/products/primary-antibodies/il1-receptor-i-il-1r-1-antibody-epr22198-36-ab229051'>ab229051</a>) at 1/1000 dilution

Lane 1:

Wild-type mouse spleen tissue lysate (female) at 20 µg

Lane 2:

IL1r1 knockout mouse spleen tissue lysate (female) at 20 µg

Lane 3:

Wild-type mouse spleen tissue lysate (male) at 20 µg

Lane 4:

IL1r1 knockout mouse spleen tissue lysate (male) at 20 µg

Lane 5:

Wild-type mouse thymus tissue lysate (female) at 20 µg

Lane 6:

IL1r1 knockout mouse thymus tissue lysate (female) at 20 µg

Lane 7:

Wild-type mouse thymus tissue lysate (male) at 20 µg

Lane 8:

IL1r1 knockout mouse thymus tissue lysate (male) at 20 µg

Secondary

All lanes:

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 81 kDa,36 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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 : 12202478).

The expression of p15 INK4b is upregulated in response to TGF beta treatment (PMID : 12202478).

The identity of the bands higher than 15 kDa are unknown.

To minimize protein degradation, cells 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-p15 INK4b antibody [EPR29010-111] (<a href='/ko/products/primary-antibodies/p15-ink4b-antibody-epr29010-111-ab323851'>ab323851</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

MCF7 treated with 10ng/ml TGF beta 1 for 18 hours whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 14-15 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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-METTL14 antibody [EPR27233-258] (<a href='/ko/products/primary-antibodies/mettl14-antibody-epr27233-258-ab323866'>ab323866</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 65 kDa,36 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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-IDUA antibody antibody [EPR26926-251] (<a href='/ko/products/primary-antibodies/idua-antibody-antibody-epr26926-251-ab323701'>ab323701</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 50 µg

Lane 2:

HeLa transfected with siRNA specifically targeting IDUA whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 60-90 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : U-937, HEL, K-562

The samples are unboiled.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 15483624).

The observed band is the N-terminal LPHN2.

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

All lanes:

Western blot - Anti-N-terminal LPHN2 antibody [EPR29557-9] (<a href='/ko/products/primary-antibodies/n-terminal-lphn2-antibody-epr29557-9-ab324484'>ab324484</a>) at 1/1000 dilution

Lane 1:

SK-N-SH (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Lane 3:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HuTu 80 (human Small intestine; Duodenum epithelial) whole cell lysate at 20 µg

Lane 5:

HEL (human erythroleukemia erythroblast) whole cell lysate at 20 µg

Lane 6:

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

Lane 7:

NIH:OVCAR-3 (human ovary adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 120 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

The bands beneath the target band (151 kDa) are likely to be degraded target fragments.

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

Exposure time : Lane 1-2 : 26 seconds; Lane 3-6 : 180 seconds

All lanes:

Western blot - Anti-EDC4 antibody [EPR28929-24] (<a href='/ko/products/primary-antibodies/edc4-antibody-epr28929-24-ab321984'>ab321984</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

SK-OV-3 (human ovarian cancer epithelial cell) whole cell lysate at 20 µg

Lane 3:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MEF (mouse embryo fibroblast) whole cell lysate at 20 µg

Lane 5:

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

Lane 6:

Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 151 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

In Western blot, ab324102 was shown to bind specifically to SETD2 (KMT3A / HYPB / HIF-1). Target of interest was observed at 120-350 kDa in wild-type Hela cell lysates (lane 1) with no signal observed at this size in SETD2 (KMT3A / HYPB / HIF-1) knockout cell line (lane 2) (lane 2, knockout cell line ab257273).

To minimize protein degradation, cells 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-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-KMT3A / HYPB / HIF-1 antibody [EPR28172-44] (<a href='/ko/products/primary-antibodies/kmt3a-hypb-hif-1-antibody-epr28172-44-ab324102'>ab324102</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

SETD2 (KMT3A / HYPB / HIF-1) knockout HeLa cell lysate knockout HeLa whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 120-350 kDa,124 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Samples are non-boiled as boiling may cause protein aggregation.

Lanes 1-4 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 and lane 5 is incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

The identity of the bands below 25 kDa in lanes 4-5 are unknown.

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

Exposure time : Lanes 1-4 : 5 seconds; Lane 5 : 26 seconds

All lanes:

Western blot - Anti-GPR89a antibody [EPR29290-504] (<a href='/ko/products/primary-antibodies/gpr89a-antibody-epr29290-504-ab325319'>ab325319</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 GPR89a whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

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

Lane 5:

Human cerebellum tissue lysate at 20 µg

Secondary

Lanes 1 - 4:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Lane 5:

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

Observed band size: 35 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Negative control : THP-1

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 12202478).

The identity of the bands higher than 15 kDa in lane 1 are unknown.

To minimize protein degradation, cells in lanes 1-2 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-p15 INK4b antibody [EPR29010-111] (<a href='/ko/products/primary-antibodies/p15-ink4b-antibody-epr29010-111-ab323851'>ab323851</a>) at 1/1000 dilution

Lane 1:

HaCaT (human skin keratinocyte) whole cell lysate at 20 µg

Lane 2:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 3:

Calu-1 (human lung epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 14-15 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

The identity of the bands at 37 kDa and 22 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-MAP3K20 (phospho S7) antibody [EPR28256-30] (<a href='/ko/products/primary-antibodies/map3k20-phospho-s7-antibody-epr28256-30-ab324636'>ab324636</a>) at 1/1000 dilution

Lane 1:

Untreated Saos-2 (human osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Saos-2 treated with 0.1 μM Nilotinib for 2 h whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 76 kDa,36 kDa

false

Exposure time: 62s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

In Western blot, ab323708 was shown to bind specifically to Dnmt3a. Target of interest was observed at 130 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in DNMT3A knockout cell line (lane 2).

The identity of the bands between 25 kDa and 37 kDa (in lanes 1-3) are unknown.

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

Exposure time : Lanes 1-2, 4-5 : 180 seconds, lane 3 : 48 seconds

All lanes:

Western blot - Anti-Dnmt3a antibody [EPR29184-89] (<a href='/ko/products/primary-antibodies/dnmt3a-antibody-epr29184-89-ab323708'>ab323708</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:

DNMT3A knockout HAP1 whole cell lysate at 20 µg

Lane 3:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

P815 (mouse mastocytoma mast Cell) whole cell lysate at 20 µg

Lane 5:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 130 kDa,36 kDa

false

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

The identity of the bands at 37 kDa and 22 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-MAP3K20 (phospho S7) antibody [EPR28256-30] (<a href='/ko/products/primary-antibodies/map3k20-phospho-s7-antibody-epr28256-30-ab324636'>ab324636</a>) at 1/1000 dilution

Lane 1:

Untreated U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate (untreated membrane) at 50 µg

Lane 2:

U-2 OS treated with 0.1 μM Nilotinib for 2 h whole cell lysate (untreated membrane) at 50 µg

Lane 3:

Untreated U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 76 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

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

In Western blot, Anti-VE Cadherin antibody [EPR27436-55] (ab313633) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-VE Cadherin (phospho Y685) antibody [EPR27437-539] (<a href='/ko/products/primary-antibodies/ve-cadherin-phospho-y685-antibody-epr27437-539-ab325018'>ab325018</a>) at 1/1000 dilution

Lane 1:

Untreated HUVEC (human umbilical vein endothelial cell) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

HUVEC treated with 100 μM Pervanadate for 10 minutes whole cell lysate (untreated membrane) at 20 µg

Lane 3:

Untreated HUVEC whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 4:

HUVEC treated with 100 μM Pervanadate for 10 minutes whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 150 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

The bands beneath the target band (151 kDa) are likely to be degraded target fragments.

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

All lanes:

Western blot - Anti-EDC4 antibody [EPR28929-24] (<a href='/ko/products/primary-antibodies/edc4-antibody-epr28929-24-ab321984'>ab321984</a>) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 151 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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='/ko/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) (ab97051) at 1/20000 dilution

Observed band size: 18 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : Daudi.

The identity of the lower MW band at approximately 25 kDa is unknown.

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

All lanes:

Western blot - Anti-Shootin-1 antibody [EPR29089-44] (<a href='/ko/products/primary-antibodies/shootin-1-antibody-epr29089-44-ab322653'>ab322653</a>) at 1/1000 dilution

Lane 1:

NCI-H69 (human lung small cell carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

SK-OV-3 (human ovarian cancer epithelial cell) whole cell lysate at 20 µg

Lane 3:

Daudi (human Burkitts lymphoma lymphoblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 53 kDa,76 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Negative control : Daudi (PMID : 35316216).

Low expression : A549 (PMID : 30250195).

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

All lanes:

Western blot - Anti-PHGDH/Malate dehydrogenase antibody [EPR28634-1] (<a href='/ko/products/primary-antibodies/phgdh-malate-dehydrogenase-antibody-epr28634-1-ab324530'>ab324530</a>) at 1/1000 dilution

Lane 1:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 6:

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

Lane 7:

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

Lane 8:

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

Lane 9:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 57 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

The identity of the bands at around 30 kDa & 25 kDa (Lane 1&3, respectively) are unknown.

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

All lanes:

Western blot - Anti-ACSL3 antibody [EPR29199-39] (<a href='/ko/products/primary-antibodies/acsl3-antibody-epr29199-39-ab321811'>ab321811</a>) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human testis tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse testis tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 80 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

The identity of the lower MW band at approximately 15 kDa is unknown.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) 1 : 100000 (15Kda).

Exposure time : Lane 1 : 180 seconds, lane 2 : 26 seconds

All lanes:

Western blot - Anti-Histone H2A (ubiquityl K119) antibody [EPR28331-576] (<a href='/ko/products/primary-antibodies/histone-h2a-ubiquityl-k119-antibody-epr28331-576-ab325796'>ab325796</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) histone fraction at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 22 kDa,15 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : brain.

The molecular weight is consistent with what have been described in literature (PMID : 12582180; 16754877).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

Exposure time : Lanes 1, 3 and 4 : 180 seconds; Lane 2 : 37 seconds

All lanes:

Western blot - Anti-MUC2 antibody [RM1281] (<a href='/ko/products/primary-antibodies/muc2-antibody-rm1281-ab322964'>ab322964</a>) at 1/1000 dilution

Lane 1:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 2:

Mouse colon tissue lysate at 20 µg

Lane 3:

Mouse small intestine tissue lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 140-170 kDa,124 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Blocking and diluting buffer and concentration : QD20250827IMG15.

Low expression control : LNCaP(PMID : 15833880), 22Rv1(PMID : 16102578).

Performed under reducing conditions.

In Western blot, ab324683 was shown to bind specifically to JAK1. Target of interest was observed at 133 kDa in wild-type HAP1 cell lysates (lane 7) with no signal observed at this size in JAK1 knockout cell line (lane 8).

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

Exposure time : Lane 1-6 : 92 seconds Lane 7, 8 : 180 seconds

All lanes:

Western blot - Anti-JAK1 antibody [EPR24649-58] (<a href='/ko/products/primary-antibodies/jak1-antibody-epr24649-58-ab324683'>ab324683</a>) at 1/1000 dilution

Lane 1:

HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate at 50 µg

Lane 2:

DU 145 (human prostate carcinoma epithelial cell) whole cell lysate at 50 µg

Lane 3:

LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 50 µg

Lane 4:

22Rv1 (human prostate carcinoma epithelial cell) whole cell lysate at 50 µg

Lane 5:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 50 µg

Lane 6:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 7:

parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 50 µg

Lane 8:

JAK1 KO HAP1 whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 133 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 22895706).

All lanes:

Western blot - Anti-TMEM106B antibody [EPR27525-70] (<a href='/ko/products/primary-antibodies/tmem106b-antibody-epr27525-70-ab321970'>ab321970</a>) at 1/1000 dilution

Lane 1:

IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

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

Lane 4:

Mouse testis tissue lysate at 20 µg

Lane 5:

Mouse kidney tissue lysate at 20 µg

Lane 6:

Mouse pancreas tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 40 kDa,80 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Samples are non-boiled as boiling may cause protein aggregation.

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

All lanes:

Western blot - Anti-GPR89a antibody [EPR29290-504] (<a href='/ko/products/primary-antibodies/gpr89a-antibody-epr29290-504-ab325319'>ab325319</a>) at 1/1000 dilution

Lanes 1 - 2:

T-47D (human ductal breast epithelial tumor epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lanes 1 - 2:

T-47D transfected with siRNA specifically targeting GPR89a whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 35 kDa,36 kDa

false

Exposure time: 62s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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-METTL14 antibody [EPR27233-258] (<a href='/ko/products/primary-antibodies/mettl14-antibody-epr27233-258-ab323866'>ab323866</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 METTL14 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 65 kDa,36 kDa

false

Exposure time: 48s

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Total protein Anti-SQSTM1 antibody [EPR18351] (ab207305) (1 : 1000).

All lanes:

Western blot - Anti-cleaved SQSTM1 / p62 at D329 (N-terminal) antibody [EPR29289-46] (<a href='/ko/products/primary-antibodies/cleaved-sqstm1-p62-at-d329-n-terminal-antibody-epr29289-46-ab325681'>ab325681</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa treated with 10ng/ml TNF alpha and 1uM IKK-16 for 4 hours whole cell lysate at 20 µg

Lane 3:

HeLa treated with 10ng/ml TNF alpha, 1uM IKK-16 and 20uM Z-VAD-FMK for 4 hours whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 40 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

To minimize protein degradation, the fresh lysate was 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-Sall4 antibody [EPR27142-83] (<a href='/ko/products/primary-antibodies/sall4-antibody-epr27142-83-ab324483'>ab324483</a>) at 1/1000 dilution

Lane 1:

NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate (fresh lysate) at 20 µg

Lane 2:

NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate (frozen lysate) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 165 kDa,36 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : Jurkat, A549

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

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

The identity of the lower MW bands below 107 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Mineralocorticoid Receptor antibody [EPR29519-576] (<a href='/ko/products/primary-antibodies/mineralocorticoid-receptor-antibody-epr29519-576-ab325523'>ab325523</a>) at 1/1000 dilution

Lane 1:

U-2 OS (human bone osteosarcoma epithelial cell) fresh whole cell lysate at 48 µg

Lane 2:

22Rv1 (human prostate carcinoma epithelial cell) fresh whole cell lysate at 48 µg

Lane 3:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) fresh whole cell lysate at 48 µg

Lane 4:

A549 (human lung carcinoma epithelial cell) fresh whole cell lysate at 48 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 107 kDa,36 kDa

true

Exposure time: 59s

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-GSDMC antibody [EPR24215-8] (<a href='/ko/products/primary-antibodies/gsdmc-antibody-epr24215-8-ab325524'>ab325524</a>) at 1/1000 dilution

Lane 1:

HCC1937 (human mammary gland epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HCC1937 transfected with siRNA specifically targeting GSDMC whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 55 kDa,36 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : MCF7.

The lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, anti-TFEB antibody (ab267351) staining at 1/1000 dilution. Anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-TFEB (phospho S211) antibody [EPR27960-64] (<a href='/ko/products/primary-antibodies/tfeb-phospho-s211-antibody-epr27960-64-ab310331'>ab310331</a>) at 1/1000 dilution

Lane 1:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

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

Lane 3:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate (phosphatase treated membrane) at 20 µg

Lane 4:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 70 kDa

false

Exposure time: 81s

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Blocking and dilution buffer and concentration : 5% NFDM /TBST.
ab181602 was used as a GAPDH loading control. ab213204 was used at 1/5000 dilution.

All lanes:

Western blot - Anti-Cytokeratin 14 + Cytokeratin 16 antibody [EP1615Y] (<a href='/ko/products/primary-antibodies/cytokeratin-16-k16-antibody-ep1615y-ab76416'>ab76416</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a His tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a full length human CK-16 expression vector containing a His-tag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a full length human CK-14 expression vector containing a His-tag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 50 kDa

false

Exposure time: 40s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

In Western blot, ab325523 was shown to bind specifically to Mineralocorticoid Receptor. Target of interest was observed at 107 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in Mineralocorticoid Receptor knockout cell line (lane 2, knockout cell line ab265540 / knockout cell lysate ab257279).

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

The identity of the lower MW bands below 107 kDa are unknown.

All lanes:

Western blot - Anti-Mineralocorticoid Receptor antibody [EPR29519-576] (<a href='/ko/products/primary-antibodies/mineralocorticoid-receptor-antibody-epr29519-576-ab325523'>ab325523</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 48 µg

Lane 2:

Mineralocorticoid Receptor knockout HeLa whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 107 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

In Western blot, ab325498 was shown to bind specifically to ZIP8/SLC39A8. Target of interest was observed at 150 kDa in wild-type HCT 166 cell lysates (lane 4) with no signal observed at this size in SLC39A8 knockout cell line (lane 5). The lysates were kindly provided by the RESOLUTE consortium.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 31261654).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-ZIP8/SLC39A8 antibody [EPR29194-220] (<a href='/ko/products/primary-antibodies/zip8-slc39a8-antibody-epr29194-220-ab325498'>ab325498</a>) at 1/1000 dilution

Lane 1:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 80 µg

Lane 2:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 80 µg

Lane 3:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 80 µg

Lane 4:

Wilde-type HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 80 µg

Lane 5:

SLC39A8 knockout HCT 116 whole cell lysate at 80 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 50 kDa,150 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Blocking and diluting buffer and concentration : 5% NFDM /TBST
ab181602 was used as a GAPDH loading control. ab213204 was used at 1/5000 dilution.

All lanes:

Western blot - Anti-Cytokeratin 14 + Cytokeratin 16 antibody [EPR13504] (<a href='/ko/products/primary-antibodies/cytokeratin-16-k16-antibody-epr13504-ab181055'>ab181055</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a His tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a full length human CK-16 expression vector containing a His-tag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a full length human CK-14 expression vector containing a His-tag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 50 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : 293T (PMID : 32299851).

The expression of GSDMB is upregulated in response to IFN gamma treatment (PMID : 32299851).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-GSDMB (C-term) antibody [EPR24636-120] (<a href='/ko/products/primary-antibodies/gsdmb-c-term-antibody-epr24636-120-ab325491'>ab325491</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

Untreated HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HT-29 treated with 10ng/ml IFN gamma for 48 hours, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 45 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : A431, HaCaT

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-GSDMC antibody [EPR24215-8] (<a href='/ko/products/primary-antibodies/gsdmc-antibody-epr24215-8-ab325524'>ab325524</a>) at 1/1000 dilution

Lane 1:

HCC1937 (human mammary gland epithelial cell) whole cell lysate at 20 µg

Lane 2:

BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

HaCaT (human skin keratinocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 55 kDa,36 kDa

false

Exposure time: 180s