AB327419

Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40]

RabMAb
Recombinant
20ul selling size
KO Validated
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Rabbit Recombinant Monoclonal ATP citrate lyase phospho S455 antibody. Suitable for WB, IHC-P and reacts with Transfected cell lysate - Human, Human, Mouse samples.

別名を表示する

ATP-citrate synthase, ATP-citrate (pro-S-)-lyase, Citrate cleavage enzyme, ACL

13 Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

Immunohistochemical analysis of paraffin-embedded
(A) Untreated Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell)
(B) Wild-type HAP1 treated with 100ng/ml Calyculin A for 30min
(C) ACLY knockout HAP1
(D) ACLY knockout HAP1 treated with 100ng/ml Calyculin A for 30min
(E) Untreated Wild-type HAP1 (alkaline phosphatase treated)
(F) Wild-type HAP1 treated with 100ng/ml Calyculin A for 30min (alkaline phosphatase treated)
(G) ACLY knockout HAP1(alkaline phosphatase treated)
(H) ACLY knockout HAP1 treated with 100ng/ml Calyculin A for 30min (alkaline phosphatase treated)
labeling with ab327419 at 1/20000 (0.026 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) Untreated Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) and (B) Wild-type HAP1 treated with 100ng/ml Calyculin A for 30min, no signal in (C) ACLY knockout HAP1, (D) ACLY knockout HAP1 treated with 100ng/ml Calyculin A for 30min, (E) Untreated Wild-type HAP1 (alkaline phosphatase treated), (F) Wild-type HAP1 treated with 100ng/ml Calyculin A for 30min (alkaline phosphatase treated), (G) ACLY knockout HAP1(alkaline phosphatase treated), (H) ACLY knockout HAP1 treated with 100ng/ml Calyculin A for 30min (alkaline phosphatase treated).

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling with ab327419 at 1/2000 (0.259 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) human breast cancer without alkaline phosphatase treatment; No signal was detected in (B) human breast cancer with alkaline phosphatase treatment.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling with ab327419 at 1/2000 (0.259 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) human lung without alkaline phosphatase treatment; No signal was detected in (B) human lung with alkaline phosphatase treatment.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling with ab327419 at 1/2000 (0.259 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) human colon without alkaline phosphatase treatment; No signal was detected in (B) human colon with alkaline phosphatase treatment.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling with ab327419 at 1/2000 (0.259 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) mouse lung without alkaline phosphatase treatment; No signal was detected in (B) mouse lung with alkaline phosphatase treatment.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

Immunohistochemical analysis of paraffin-embedded
(A) Untreated NIH/3T3 (mouse embryonic fibroblast)
(B) NIH/3T3 treated with 50ng/ml Platelet-Derived Growth Factor-BB for 60 min
(C) Untreated NIH/3T3 (alkaline phosphatase treated)
(D) NIH/3T3 treated with 50ng/ml Platelet-Derived Growth Factor-BB for 60 min (alkaline phosphatase treated) tissue labeling with ab327419 at 1/20000 (0.026 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Weak staining in (A) Untreated NIH/3T3 (mouse embryonic fibroblast), with increased expression of ACLY in (B) NIH/3T3 treated with 50ng/ml Platelet-Derived Growth Factor-BB for 60 min, and no signal in (C) Untreated NIH/3T3 (alkaline phosphatase treated), (D) NIH/3T3 treated with 50ng/ml Platelet-Derived Growth Factor-BB for 60 min (alkaline phosphatase treated).

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling with ab327419 at 1/2000 (0.259 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) mouse kidney without alkaline phosphatase treatment; No signal was detected in (B) mouse kidney with alkaline phosphatase treatment.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

Immunohistochemical analysis of paraffin-embedded Mouse breast cancer tissue labeling with ab327419 at 1/2000 (0.259 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) mouse breast cancer without alkaline phosphatase treatment; No signal was detected in (B) mouse breast cancer with alkaline phosphatase treatment.

The primary antibody was incubated 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

Lab

Western blot - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

Blocking buffer and concentration : 1% BSA/TBST
Diluting buffer and concentration : 5% NFDM/TBST

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa);Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(125KDa).

All lanes:

Western blot - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (ab327419) at 1/1000 dilution

Lane 1:

293T cells transfected with a human wild-type ACLY expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human ACLY(S455A mutant) 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='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 125 kDa,36 kDa

false

Exposure time: 1s

Lab

Western blot - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

Blocking buffer and concentration : 1% BSA/TBST
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-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (ab327419) at 1/1000 dilution

Lane 1:

Human colon tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse colon tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 125 kDa,36 kDa

false

Exposure time: 180s

Lab

Western blot - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

Blocking buffer and concentration : 1% BSA/TBST
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-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (ab327419) at 1/1000 dilution

Lane 1:

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

Lane 2:

HepG2 treated with 150nM Insulin for 15 min whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 125 kDa,36 kDa

false

Exposure time: 125s

Lab

Western blot - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

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-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (ab327419) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

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

Lane 4:

NIH/3T3 treated with 50ng/ml Platelet-Derived Growth Factor-BB for 60 min whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 125 kDa,36 kDa

false

Exposure time: 125s

Lab

Western blot - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (AB327419)

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

In Western blot, ab327419 was shown to bind specifically to ATP citrate lyase (phospho S455). Target of interest was observed at 125 kD wild-type HAP1 cell lysates (lanes 1-2) with no signal observed at this size in ACLY knockout cell line (lanes 3-4) .

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

All lanes:

Western blot - Anti-ATP citrate lyase (phospho S455) antibody [EPR31107-40] (ab327419) at 1/1000 dilution

Lane 1:

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

Lane 2:

Wild-type HAP1 treated with 100ng/ml Calyculin A for 30min whole cell lysate (untreated membrane) at 20 µg

Lane 3:

ACLY knockout HAP1 whole cell lysate (untreated membrane) at 20 µg

Lane 4:

ACLY knockout HAP1 treated with 100ng/ml Calyculin A for 30min whole cell lysate (untreated membrane) at 20 µg

Lane 5:

Untreated Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 6:

Wild-type HAP1 treated with 100ng/ml Calyculin A for 30min whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 7:

ACLY knockout HAP1 whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 8:

ACLY knockout HAP1 treated with 100ng/ml Calyculin A for 30min whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 125 kDa,36 kDa

false

Exposure time: 3s

Key facts

宿主種

Rabbit

クローン性

Monoclonal

クローン番号

EPR31107-40

アイソタイプ

IgG

キャリアフリー

交差種

Human, Mouse

アプリケーション

WB, IHC-P

applications

免疫原

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/2000 - 1/20000", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol." }, "Mouse": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/2000 - 1/20000", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol." }, "Transfected cell lysate - Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }

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製品の詳細

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

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出荷温度及び保存条件

製品の状態

Liquid

精製方法

Affinity purification Protein A

バッファー組成

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

出荷温度

Conditional Ambient

短期保存期間

1-2 weeks

短期保存温度

+4°C

長期保存温度

-20°C

分注に関する情報

Upon delivery aliquot

保管に関する情報

Avoid freeze / thaw cycle

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製品プロトコール

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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ターゲットの情報

Catalyzes the cleavage of citrate into oxaloacetate and acetyl-CoA, the latter serving as common substrate in multiple biochemical reactions in protein, carbohydrate and lipid metabolism.

See full target information ACLY pS455

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Abcam product promise

当社は、高品質な試薬を通じてお客様の研究を力強くサポートすることをお約束いたします。ご使用いただく各段階で、常にお客様をサポートできる体制を整えております。万が一、製品が期待通りに機能しない場合は、「Abcam Product Promise」による当社保証制度に基づき、安心してご利用いただけます。

保証に関する詳細については利用規約をご確認ください。

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