Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)

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 - BSA and Azide free
  • Suitable for: IP, WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free
    See all FHL1 primary antibodies

  • Description

    Rabbit monoclonal [EPR22842-95] to FHL1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, WB, IHC-P, ICC/IFmore 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 and HEK293T cell lysates. NCI-H1299 whole cell lysate. 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

    Ab256551 is the carrier-free version of ab255828. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab256551 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Properties

Images

  • Western blot - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)
    Western blot - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)
    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?



    This data was developed using ab255828, the same antibody clone in a different buffer formulation.

    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] - BSA and Azide free (ab256551)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255828).

  • Immunoprecipitation - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)
    Immunoprecipitation - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255828).

  • Immunoprecipitation - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)
    Immunoprecipitation - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255828).

  • Immunocytochemistry/ Immunofluorescence - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)
    Immunocytochemistry/ Immunofluorescence - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255828).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255828).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255828).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255828).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255828).

  • Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)
    Anti-FHL1 antibody [EPR22842-95] - BSA and Azide free (ab256551)

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