Name: Anti-PFKFB3 antibody [EPR12594]
Brand: RabMAb
SKU: ab181861
Rating: 5 (6 reviews)

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Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR12594] to PFKFB3
Suitable for: Flow Cyt (Intra), WB, IP, ICC/IF, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

产品名称

Anti-PFKFB3抗体[EPR12594]
参阅全部 PFKFB3 一抗
描述

兔单克隆抗体[EPR12594] to PFKFB3
宿主

Rabbit
经测试应用

适用于: Flow Cyt (Intra), WB, IP, ICC/IF, IHC-Pmore details
种属反应性

与反应: Mouse, Rat, Human
免疫原

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
阳性对照
- WB: Jurkat and HeLa whole cell lysate (ab150035); Human melanoma tissue; HeLa and A431 cells, Mouse skin tissue lysate, Rat breast tissue lysate, AR42 and L6 whole cell lysates, HAP1 whole cell lysate, AR42J (rat pancreatic tumor epithelial cell) whole cell lysate, IP: Mouse skin tissue lysate, AR42J, whole cell lysate ICC: HeLa, A431 cells IHC: human melanoma tissue Flow: A431 (human epidermoid carcinoma) cells,
常规说明
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

形式

Liquid
存放说明

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
存储溶液

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
纯度

Protein A purified
克隆

单克隆
克隆编号

EPR12594
同种型

IgG
研究领域

The Abpromise guarantee

Abpromise™承诺保证使用ab181861于以下的经测试应用

“应用说明”部分下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用	Ab评论	说明
Flow Cyt (Intra)		Use at an assay dependent concentration.
WB	(5)	1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 60 kDa).
IP		1/50.
ICC/IF		1/100.
IHC-P		1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

说明
Flow Cyt (Intra) Use at an assay dependent concentration.
WB 1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 60 kDa).
IP 1/50.
ICC/IF 1/100.
IHC-P 1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

功能

Synthesis and degradation of fructose 2,6-bisphosphate.
组织特异性

Ubiquitous.
序列相似性

In the C-terminal section; belongs to the phosphoglycerate mutase family.
翻译后修饰

Phosphorylation by AMPK stimulates activity.
Target information above from: UniProt accession Q16875 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
数据库链接
- Entrez Gene: 5209 Human
- Entrez Gene: 170768 Mouse
- Entrez Gene: 117276 Rat
- Omim: 605319 Human
- SwissProt: Q16875 Human
- SwissProt: A2AUP1 Mouse
- SwissProt: O35552 Rat
- Unigene: 195471 Human
别名
- 6 phosphofructo 2 kinase/ fructose 2,6 bisphosphatase antibody
- 6 phosphofructo 2 kinase/fructose 2,6 biphosphatase 3 antibody
- 6-bisphosphatase antibody
- 6-P2ase 3 antibody
- 6-P2ASE brain/placenta-type isozyme antibody
- 6PF 2 K/Fru 2,6 P2ASE brain/placenta type isozyme antibody
- 6PF 2-K/Fru 2,6 P2ase 3 antibody
- 6PF-2-K/Fru-2 antibody
- F263_HUMAN antibody
- fructose 6 phosphate,2 kinase/fructose 2, 6 bisphosphatase antibody
- Fructose-2 antibody
- Inducible 6 phosphofructo 2 kinase/fructose 2,6 bisphosphatase antibody
- iPFK 2 antibody
- iPFK-2 antibody
- IPFK2 antibody
- PFK/FBPase 3 antibody
- PFK2 antibody
- PFKFB3 antibody
- Renal carcinoma antigen NY REN 56 antibody
- Renal carcinoma antigen NY-REN-56 antibody
- uPFK antibody
see all

Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: PFKFB3 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: Jurkat whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab181861 observed at 60 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab181861 was shown to specifically react with PFKFB3 in wild-type HAP1 cells as signal was lost in PFKFB3 knockout cells. Wild-type and PFKFB3 knockout samples were subjected to SDS-PAGE. Ab181861 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] (ab181861)

Immunofluorescent analysis of acetone-fixed HeLa cells labeling PFKFB3 with ab181861 at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor®555) at 1/200 dilution. Counter stained with Dapi (blue).
Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861)

All lanes : Anti-PFKFB3 antibody [EPR12594] (ab181861) at 1/20000 dilution

Lane 1 : Jurkat cell lysate
Lane 2 : HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 60 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861)

All lanes : Anti-PFKFB3 antibody [EPR12594] (ab181861) at 1/1000 dilution

Lane 1 : A431 (human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 2 : Mouse skin tissue lysate
Lane 3 : Rat breast tissue lysate
Lane 4 : AR42J (rat pancreatic tumor epithelial cell) whole cell lysate
Lane 5 : L6 (rat skeletal muscle myoblast) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 60 kDa
Observed band size: 60 kDa

Exposure time: 15 seconds

Blocking buffer: 5% NFDM/TBST

Exposure time: 15 seconds

This blot was developed using a high sensitivity ECL substrate.
Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861)

All lanes : Anti-PFKFB3 antibody [EPR12594] (ab181861) at 1/1000 dilution

Lane 1 : A431 (human epidermoid carcinoma epithelial cell), whole cell lysate
Lane 2 : bEnd.3 (mouse brain endothelial cell), whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
Lane 4 : 4T1 (mouse mammary gland carcinoma epithelial cell), whole cell lysate
Lane 5 : Undifferentiated 3T3-L1 (mouse embryonic fibroblast), whole cell lysate
Lane 6 : 3T3-L1 (mouse embryonic fibroblast) differentiated into adipocyte-like cells, whole cell lysate
Lane 7 : C2C12 (mouse myoblast), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 60 kDa
Observed band size: 60 kDa

Exposure time: Lane 1-2: 26 seconds, Lane 3-7: 48 seconds

Blocking buffer and concentration: 5% NFDM/TBST

Lane 3-7 were developed using a high sensitivity ECL substrate.
The expression level of PFKFB3 is upregulated during 3T3-L1 differentiation (PMID: 16306349).
The band at approximately 110 kDa is likely to be PFKFB3 dimer (PMID: 31889092).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PFKFB3 antibody [EPR12594] (ab181861)

Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling PFKFB3 with ab181861 at 1/50 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-PFKFB3 antibody [EPR12594] (ab181861)

Intracellular Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labeling PFKFB3 with purified ab181861 at 1/210 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.
Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (ab181861)

Western blot analysis of PFKFB3 in HeLa cell lysate immunoprecipitated using ab181861 at 1/50 dilution (Lane 1). Lane 2: Negative control.
Secondary antibody: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution.
Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (ab181861)

PFKFB3 was immunoprecipitated from 0.35 mg of Mouse skin tissue lysate with ab181861 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab181861 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: Mouse skin tissue lysate 10 μg (Input).

Lane 2: ab181861 IP in Mouse skin tissue lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab181861 in Mouse skin tissue lysate

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

Exposure time: 180 seconds.

This blot was developed using a high sensitivity ECL substrate.
Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (ab181861)

PFKFB3 was immunoprecipitated from 0.35 mg of AR42J (rat pancreatic tumor epithelial cell) whole cell lysate with ab181861 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab181861 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: AR42J (rat pancreatic tumor epithelial cell) whole cell lysate 10 μg (Input).

Lane 2: ab181861 IP in AR42J whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab181861 in AR42J whole cell lysate

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

Exposure time: 180 seconds.
Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] (ab181861)

Immunofluorescent analysis of 4% paraformaldehyde-fixed A431 cells labeling PFKFB3 with ab181861 at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor®555) at 1/200 dilution. Counter stained with Dapi (blue).
Anti-PFKFB3 antibody [EPR12594] (ab181861)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

发表研究结果有使用 ab181861？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab181861 被引用在 73 文献中.

Mela V et al. Acute Stress, Induced by IFNγ + Aβ, and Chronic Stress, Induced by Age, Affect Microglia in a Sex-Specific Manner. Mol Neurobiol 60:3044-3053 (2023). PubMed: 36781739
Su W et al. Hexokinase 2-mediated glycolysis supports inflammatory responses to Porphyromonas gingivalis in gingival fibroblasts. BMC Oral Health 23:103 (2023). PubMed: 36793034
Paladines N et al. Metabolic reprogramming through mitochondrial biogenesis drives adenosine anti-inflammatory effects: new mechanism controlling gingival fibroblast hyper-inflammatory state. Front Immunol 14:1148216 (2023). PubMed: 37350964
Huang X et al. PFKFB3 downregulation aggravates Angiotensin II-induced podocyte detachment. Ren Fail 45:2230318 (2023). PubMed: 37427767
Yang Q et al. PFKFB3-Mediated Glycolysis Boosts Fibroblast Activation and Subsequent Kidney Fibrosis. Cells 12:N/A (2023). PubMed: 37626891

View all Publications for this product

客户评价及客户问答

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1-6 of 6 Abreviews or Q&A

Application

IHC - Wholemount

Sample

Mouse Tissue (Lung)

Specification

Lung

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Ming Jen Hsu

Verified customer

提交于 Aug 18 2021

Application

Western blot

Sample

Mouse Tissue lysate - whole (brain)

Gel Running Conditions

Reduced Denaturing (TEO TRICINE 4-20% gel (ab119209))

Loading amount

20 µg

Specification

brain

Blocking step

BSA as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Yonatan Feuermann

Verified customer

提交于 Apr 12 2021

Application

Western blot

Sample

Mouse Cell lysate - whole cell (Murine breast cancer cells)

Gel Running Conditions

Reduced Denaturing (10% gel)

Loading amount

5 µg

Specification

Murine breast cancer cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Alyssa La Belle

Verified customer

提交于 May 21 2019

Application

Western blot

Sample

Cow Cell lysate - whole cell (VVEC)

Gel Running Conditions

Reduced Denaturing (10)

Loading amount

20 µg

Treatment

100 uM ATP 0, 5 min, 30 min

Specification

VVEC

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jun 03 2016

Application

Western blot

Sample

Mouse Cell lysate - whole cell (primary alveolar type II cells)

Gel Running Conditions

Non-reduced Denaturing (10%)

Loading amount

20 µg

Specification

primary alveolar type II cells

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Nov 02 2015

Application

Western blot

Sample

Human Cell lysate - whole cell (A549 cells)

Gel Running Conditions

Non-reduced Denaturing (10)

Loading amount

20 µg

Specification

A549 cells

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Christine Vohwinkel

Verified customer

提交于 Oct 29 2015

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Key features and details

Related conjugates and formulations

概述

产品名称

描述

宿主

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Positive Controls

应用

The Abpromise guarantee

靶标

功能

组织特异性

序列相似性

翻译后修饰

数据库链接

别名

图片

实验方案

数据表及文件

SDS download

Datasheet download

Certificate of Compliance

文献 (73)

客户评价及客户问答

IHC - Wholemount abreview for Anti-PFKFB3 antibody [EPR12594]

Western blot abreview for Anti-PFKFB3 antibody [EPR12594]

Western blot abreview for Anti-PFKFB3 antibody [EPR12594]

Western blot abreview for Anti-PFKFB3 antibody [EPR12594]

Western blot abreview for Anti-PFKFB3 antibody [EPR12594]

Western blot abreview for Anti-PFKFB3 antibody [EPR12594]