Anti-FHL2 antibody [EPR17860-23] (ab202586)

Rabbit Recombinant Monoclonal FHL2 antibody. Suitable for IP, WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 1 publication.

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Images

Western blot - Anti-FHL2 antibody [EPR17860-23] (AB202586), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

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

Reactivity data

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Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF
Human	Tested	Tested	Tested
Mouse	Expected	Tested	Tested
Rat	Expected	Tested	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/20	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/100	Notes ab202586 works both with PFA and methanol fixation.
Species Human	Dilution info 1/100	Notes ab202586 works both with PFA and methanol fixation.

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

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Target data

See full target information FHL2

Function

May function as a molecular transmitter linking various signaling pathways to transcriptional regulation. Negatively regulates the transcriptional repressor E4F1 and may function in cell growth. Inhibits the transcriptional activity of FOXO1 and its apoptotic function by enhancing the interaction of FOXO1 with SIRT1 and FOXO1 deacetylation. Negatively regulates the calcineurin/NFAT signaling pathway in cardiomyocytes (PubMed:28717008).

Alternative names

DRAL, SLIM3, FHL2, Four and a half LIM domains protein 2, FHL-2, LIM domain protein DRAL, Skeletal muscle LIM-protein 3, SLIM-3

Recommended products

Rabbit Recombinant Monoclonal FHL2 antibody. Suitable for IP, WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 1 publication.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR17860-23
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

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

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The FHL2 protein also known as Four and a Half LIM Domains Protein 2 plays an important role in cellular mechanics. Its molecular mass is approximately 32 kDa. FHL2 is expressed in various tissues with significant presence in cardiac and skeletal muscles as well as the liver. It is involved in the regulation of gene expression by binding to other proteins and DNA through its LIM domains. This ability allows FHL2 to influence cellular structures and functions integrally.

Biological function summary

The FHL2 protein serves critical functions in muscle development and differentiation. It operates as part of a multiprotein complex that interacts with cytoskeletal components contributing importantly to muscle cell adhesion and communication. Moreover FHL2 engages in transcriptional regulation and acts as a coactivator or corepressor of specific transcription factors influencing gene expression patterns relevant to muscle function and maintenance.

Pathways

The FHL2 protein plays significant roles in Wnt and MAPK signaling pathways. In the Wnt signaling pathway FHL2 regulates the transcriptional activity of beta-catenin an essential component of the pathway involved in cell growth and differentiation. Additionally FHL2 contributes to the MAPK signaling pathway by influencing interactions between upstream regulators and downstream targets modulating cellular responses to external stimuli. Through these pathways FHL2 interacts with various proteins including GATA4 and TEF-1 which are vital for its regulatory functions.

Associated diseases and disorders

FHL2 has associations with certain cancers like prostate cancer and cardiomyopathy. In prostate cancer FHL2 overexpression leads to increased tumor progression and is often linked to androgen receptors promoting malignant transformation. In cardiomyopathy alterations in FHL2 expression affect heart muscle integrity and function sometimes in conjunction with interactions involving desmin an important structural protein. Understanding FHL2's connections to these diseases offers insights into potential therapeutic targets.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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11 product images

Western blot - Anti-FHL2 antibody [EPR17860-23] (ab202586)
Lanes 1 - 3: Merged signal (red and green). Green - ab202586 observed at 32 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab202586 was shown to react with FHL2 in wild-type U-2 OS cells in western blot with loss of signal observed in FHL2 knockout sample. Wild-type and FHL2 knockout U-2 OS cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab202586 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-FHL2 antibody [EPR17860-23] (ab202586) at 1/1000 dilution
Lane 1: Wild-type U-2 OS cell lysate at 40 µg
Lane 2: FHL2 knockout U-2 OS cell lysate at 40 µg
Lane 3: HeLa cell lysate at 40 µg
Predicted band size: 32 kDa
Western blot - Anti-FHL2 antibody [EPR17860-23] (ab202586)
Lanes 1 - 3: Merged signal (red and green). Green - ab202586 observed at 32 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab202586 was shown to react with FHL2 in wild-type U-2 OS cells in western blot with loss of signal observed in FHL2 knockout sample. Wild-type and FHL2 knockout U-2 OS cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab202586 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-FHL2 antibody [EPR17860-23] (ab202586) at 1/1000 dilution
Lane 1: Wild-type U-2 OS cell lysate at 40 µg
Lane 2: FHL2 knockout U-2 OS cell lysate at 40 µg
Lane 2: Western blot - Human FHL2 knockout U-2 OS cell line (Human FHL2 knockout U-2 OS cell line ab262496)
Lane 3: HeLa cell lysate at 40 µg
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 32 kDa
Immunocytochemistry/ Immunofluorescence - Anti-FHL2 antibody [EPR17860-23] (ab202586)
ab202586 staining FHL2 in wild-type U2OS cells (top panel) and FHL2 knockout U2OS cells (bottom panel). The cells were fixed with 4% PFA (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab202586 at 1/100 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Western blot - Anti-FHL2 antibody [EPR17860-23] (ab202586)
** Lanes 1-3:** Merged signal (red and green). Green - ab202586 observed at 32 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 observed at 50 kDa.
ab202586 Recombinant Anti-FHL2 antibody [EPR17860-23] was shown to specifically react with FHL2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human FHL2 knockout HeLa cell line ab265475 (knockout cell lysate Human FHL2 knockout HeLa cell lysate ab257441) was used. Wild-type and FHL2 knockout samples were subjected to SDS-PAGE. ab202586 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-FHL2 antibody [EPR17860-23] (ab202586) at 1/1000 dilution
Lane 1: Wild-type HeLa lysate at 20 µg
Lane 2: FHL2 knockout HeLa lysate at 20 µg
Lane 2: Western blot - Human FHL2 knockout HeLa cell line (Human FHL2 knockout HeLa cell line ab265475)
Lane 3: K562 lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 32 kDa
Immunocytochemistry/ Immunofluorescence - Anti-FHL2 antibody [EPR17860-23] (ab202586)
ab202586 staining FHL2 in wild-type U2OS cells (top panel) and FHL2 knockout U2OS cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab202586 at 1/100 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Immunocytochemistry/ Immunofluorescence - Anti-FHL2 antibody [EPR17860-23] (ab202586)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A-673 (Human muscle Ewing's Sarcoma cell line) cells labeling FHL2 with ab202586 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining on A-673 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab202586 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/500 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/500 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-FHL2 antibody [EPR17860-23] (ab202586)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling FHL2 with ab202586 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining on A-673 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab202586 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/500 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/500 dilution.
Immunoprecipitation - Anti-FHL2 antibody [EPR17860-23] (ab202586)
FHL2 was immunoprecipitated from 1mg of SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate with ab202586 at 1/20 dilution. Western blot was performed from the immunoprecipitate using ab202586 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: SW480 whole cell lysate 10 μg (Input). Lane 2: ab202586 IP in SW480 whole cell lysate. Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab202586 in SW480 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
All lanes: Immunoprecipitation - Anti-FHL2 antibody [EPR17860-23] (ab202586)
Predicted band size: 32 kDa
Western blot - Anti-FHL2 antibody [EPR17860-23] (ab202586)
5% NFDM/TBST: Blocking and diluting buffer.
All lanes: Western blot - Anti-FHL2 antibody [EPR17860-23] (ab202586) at 1/2000 dilution
Lane 1: SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate at 10 µg
Lane 2: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg
Lane 3: HT1080 (Human fibrosarcoma cells) whole cell lysate at 10 µg
Lane 4: HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution
Predicted band size: 32 kDa
Observed band size: 32 kDa
Exposure time: 3min
Western blot - Anti-FHL2 antibody [EPR17860-23] (ab202586)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-FHL2 antibody [EPR17860-23] (ab202586) at 1/10000 dilution
All lanes: Human fetal heart lysate at 10 µg
Secondary
All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 32 kDa
Observed band size: 32 kDa
Exposure time: 30s
Western blot - Anti-FHL2 antibody [EPR17860-23] (ab202586)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-FHL2 antibody [EPR17860-23] (ab202586) at 1/10000 dilution
Lane 1: Mouse heart lysate at 10 µg
Lane 2: Mouse kidney lysate at 10 µg
Lane 3: Rat heart lysate at 10 µg
Lane 4: Rat kidney lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution
Predicted band size: 32 kDa
Observed band size: 32 kDa
Exposure time: 30s