Recombinant Anti-FHL1 antibody [EPR22842-95] (ab255828)
![Western blot - Anti-FHL1 antibody [EPR22842-95] (ab255828)](https://www.abcam.com/ps/products/255/ab255828/Images/ab255828-410218-anti-fhl1-antibody-epr22842-95-western-blot-human-knockout-lysate.jpg "Western blot - Anti-FHL1 antibody [EPR22842-95] (ab255828)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22842-95] to FHL1
- Suitable for: ICC/IF, IHC-P, IP, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-FHL1 antibody [EPR22842-95]
See all FHL1 primary antibodies -
Description
Rabbit monoclonal [EPR22842-95] to FHL1 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IHC-P, IP, WBmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: HeLa, HEK293T and NCI-H1299 whole cell lysates. Mouse, human and rat skeletal muscle lysate. Human lung, mouse kidney, mouse spleen and rat heart lysate IHC-P: Human skeletal muscle, cardiac muscle and colon tissue. Mouse lung and rat skeletal muscle tissue. ICC/IF: NCI-H1299 cells. IP: Human lung and mouse skeletal muscle lysate.
-
General notes
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
-
Form
Liquid -
Storage instructions
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. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22842-95 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab255828 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF |
1/50.
|
|
| IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
| IP |
1/30.
|
|
| WB |
1/1000. Predicted molecular weight: 36 kDa.
|
| Notes |
|---|
|
ICC/IF
1/50.
|
|
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
IP
1/30.
|
|
WB
1/1000. Predicted molecular weight: 36 kDa.
|
Target
-
Function
May have an involvement in muscle development or hypertrophy. -
Tissue specificity
Isoform 1 is highly expressed in skeletal muscle and to a lesser extent in heart, placenta, ovary, prostate, testis, small intestine, colon and spleen. Expression is barely detectable in brain, lung, liver, kidney, pancreas, thymus and peripheral blood leukocytes. Isoform 2 is expressed in brain, skeletal muscle and to a lesser extent in heart, colon, prostate and small intestine. Isoform 3 is expressed in testis, heart and skeletal muscle. -
Involvement in disease
Defects in FHL1 are the cause of X-linked dominant scapuloperoneal myopathy (SPM) [MIM:300695]. Scapuloperoneal syndrome (SPS) was initially described more than 120 years ago by Jules Broussard as 'une forme hereditaire d'atrophie musculaire progressive' beginning in the lower legs and affecting the shoulder region earlier and more severely than distal arm. The etiology of this condition remains unclear.
Defects in FHL1 are the cause of X-linked myopathy with postural muscle atrophy (XMPMA) [MIM:300696]. Myopathies are inherited muscle disorders characterized by weakness and atrophy of voluntary skeletal muscle, and several types of myopathy also show involvement of cardiac muscle. XMPMA is a distinct form of adult-onset X-linked recessive myopathy with several features in common with other myopathies, but the presentation of a pseudoathletic phenotype, scapuloperoneal weakness, and bent spine is unique and might render the clinical phenotype distinguishable from other myopathies.
Defects in FHL1 are the cause of X-linked severe early-onset reducing body myopathy (RBM) [MIM:300717]. RBM is a rare muscle disorder causing progressive muscular weakness and characteristic intracytoplasmic inclusions in myofibers. Clinical presentations of RBM have ranged from early onset fatal to childhood onset to adult onset cases.
Defects in FHL1 are the cause of X-linked childhood-onset reducing body myopathy (CO-RBM) [MIM:300718]. This disorder is allelic to severe early-onset reducing body myopathy (RBM) [MIM:300717]. -
Sequence similarities
Contains 3 LIM zinc-binding domains. -
Developmental stage
Elevated levels during postnatal muscle growth. -
Cellular localization
Cytoplasm; Cytoplasm. Nucleus and Nucleus. Cytoplasm > cytosol. Predominantly nuclear in myoblasts but is cytosolic in differentiated myotubes. - Information by UniProt
-
Database links
- Entrez Gene: 2273 Human
- Entrez Gene: 14199 Mouse
- Entrez Gene: 25177 Rat
- Omim: 300163 Human
- SwissProt: Q13642 Human
- SwissProt: P97447 Mouse
- SwissProt: Q9WUH4 Rat
- Unigene: 435369 Human
see all -
Alternative names
- bA535K18.1 antibody
- FHL 1 antibody
- FHL 1B antibody
see all
Images
-
Western blot - Anti-FHL1 antibody [EPR22842-95] (ab255828)All lanes : Anti-FHL1 antibody [EPR22842-95] (ab255828) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : FHL1 knockout HeLa cell lysate
Lane 3 : HEK293T cell lysate
Lane 4 : Daudi cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab255828 observed at 32 kDa. Red - loading control ab7291 observed at 50 kDa.
ab255828 Anti-FHL1 antibody [EPR22842-95] was shown to specifically react with FHL1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266011 (knockout cell lysate ab257952) was used. Wild-type and FHL1 knockout samples were subjected to SDS-PAGE. ab255828 and 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 (ab216773) and 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.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] (ab255828)](https://www.abcam.com/ps/products/255/ab255828/Images/ab255828-3-anti-fhl1-antibody-epr22842-95-immunohistochemistry-colon-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] (ab255828)Immunohistochemical analysis of paraffin-embedded human colon tissue labeling FHL1 with ab255828 at 1/500 dilution (1.036 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the stroma and smooth muscle in human colon is observed. The section was incubated with ab255828 for 30 mins at RT. 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
-
![Immunocytochemistry/ Immunofluorescence - Anti-FHL1 antibody [EPR22842-95] (ab255828)](https://www.abcam.com/ps/products/255/ab255828/Images/ab255828-6-anti-fhl1-antibody-epr22842-95-immunocytochemistry-ncih1299-human.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-FHL1 antibody [EPR22842-95] (ab255828)Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NCI-H1299 (Human lung carcinoma epithelial cell) cells labeling FHL1 with ab255828 at 1/50 10 µg/ml dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 2 µg/ml dilution (Green). Confocal image showing cytoplasmic and nuclear staining in NCI-H1299 cell line. Low expression control: NCI-H460 cell line (PMID: 21702045). ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 2 µg/ml dilution.
-
![Western blot - Anti-FHL1 antibody [EPR22842-95] (ab255828)](https://www.abcam.com/ps/products/255/ab255828/Images/ab255828-353726-anti-fhl1-antibody-epr22842-95-western-blot.jpg)
Western blot - Anti-FHL1 antibody [EPR22842-95] (ab255828)All lanes : Anti-FHL1 antibody [EPR22842-95] (ab255828) at 1/1000 dilution
Lane 1 : NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysate
Lane 2 : NCI-H460 (human large cell lung cancer epithelial cell) whole cell lysate
Lane 3 : Mouse skeletal muscle lysate
Lane 4 : Human skeletal muscle lysate
Lane 5 : Human lung lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lanes 4-5 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?There are five isoforms of FHL1. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 21702045).
Low expression control: NCI-H460 (PMID: 21702045).
Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
Exposure Time:
Lanes 1-2: 3 minutes;
Lane 3: 26 seconds;
Lane 4: 1 second;
Lane 5: 26 seconds.
-
![Western blot - Anti-FHL1 antibody [EPR22842-95] (ab255828)](https://www.abcam.com/ps/products/255/ab255828/Images/ab255828-353725-anti-fhl1-antibody-epr22842-95-western-blot-2.jpg)
Western blot - Anti-FHL1 antibody [EPR22842-95] (ab255828)All lanes : Anti-FHL1 antibody [EPR22842-95] (ab255828) at 1/1000 dilution
Lane 1 : Mouse kidney lysate
Lane 2 : Mouse spleen lysate
Lane 3 : Rat heart lysate
Lane 4 : Rat skeletal muscle lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 36 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?There are five isoforms of FHL1. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 21702045).
Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
Exposure Time:
Lanes 1-3: 3 minutes;
Lane 4: 3 seconds.
-
![Immunoprecipitation - Anti-FHL1 antibody [EPR22842-95] (ab255828)](https://www.abcam.com/ps/products/255/ab255828/Images/ab255828-8-anti-fhl1-antibody-epr22842-95-immunoprecipitation-skeletal-muscle-mouse.jpg)
Immunoprecipitation - Anti-FHL1 antibody [EPR22842-95] (ab255828)FHL1 was immunoprecipitated from 0.35 mg mouse skeletal muscle lysate 10µg with ab255828 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab255828 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: Mouse skeletal muscle lysate 10µg.
Lane 2: ab255828 IP in mouse skeletal muscle lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab255828 in mouse skeletal muscle lysate.
Blocking and dilution buffer and concentration/ 5% NFDM/TBST.
Exposure time/ 30 seconds.
-
![Immunoprecipitation - Anti-FHL1 antibody [EPR22842-95] (ab255828)](https://www.abcam.com/ps/products/255/ab255828/Images/ab255828-7-anti-fhl1-antibody-epr22842-95-immunoprecipitation-lung-human.jpg)
Immunoprecipitation - Anti-FHL1 antibody [EPR22842-95] (ab255828)FHL1 was immunoprecipitated from 0.35 mg human lung lysate 10µg with ab255828 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab255828 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: Human lung lysate 10µg.
Lane 2: ab255828 IP in human lung lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab255828 in human lung lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time/ 30 seconds.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] (ab255828)](https://www.abcam.com/ps/products/255/ab255828/Images/ab255828-5-anti-fhl1-antibody-epr22842-95-immunohistochemistry-skeletal-muscle-rat.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] (ab255828)Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue labeling FHL1 with ab255828 at 1/500 dilution (1.036 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on rat skeletal muscle (PMID: 27443559, 9714789) is observed. The section was incubated with ab255828 for 30 mins at RT. 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] (ab255828)](https://www.abcam.com/ps/products/255/ab255828/Images/ab255828-4-anti-fhl1-antibody-epr22842-95-immunohistochemistry-lung-mouse.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] (ab255828)Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling FHL1 with ab255828 at 1/500 dilution (1.036 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on mouse lung (PMID: 29067108) is observed. The section was incubated with ab255828 for 30 mins at RT. 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] (ab255828)](https://www.abcam.com/ps/products/255/ab255828/Images/ab255828-2-anti-fhl1-antibody-epr22842-95-immunohistochemistry-cardiac-muscle-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] (ab255828)Immunohistochemical analysis of paraffin-embedded human cardiac muscle tissue labeling FHL1 with ab255828 at 1/500 dilution (1.036 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human cardiac muscle (PMID: 9714789) is observed. The section was incubated with ab255828 for 30 mins at RT. 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] (ab255828)](https://www.abcam.com/ps/products/255/ab255828/Images/ab255828-1-anti-fhl1-antibody-epr22842-95-immunohistochemistry-skeletal-muscle-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] (ab255828)Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling FHL1 with ab255828 at 1/500 dilution (1.036 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human skeletal muscle (PMID: 27443559, 9714789) is observed. The section was incubated with ab255828 for 30 mins at RT. 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
-
![Anti-FHL1 antibody [EPR22842-95] (ab255828)](https://www.abcam.com/ps/products/255/ab255828/Images/ab255828-12-benefits-of-recombinant-antibodies.png)
Anti-FHL1 antibody [EPR22842-95] (ab255828)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (0)
ab255828 has not yet been referenced specifically in any publications.