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Recombinant

Recombinant Anti-GFAP antibody [RM1003] - BSA and Azide free (ab283862)

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Western blot - Anti-GFAP antibody [RM1003] - BSA and Azide free (ab283862)
  • Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [RM1003] - BSA and Azide free (ab283862)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [RM1003] - BSA and Azide free (ab283862)
  • Flow Cytometry (Intracellular) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
  • Western blot - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
  • Immunoprecipitation - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
  • Western blot - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
  • Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit recombinant multiclonal [RM1003] to GFAP - BSA and Azide free
  • Suitable for: WB, Flow Cyt (Intra), IHC-P, IP, IHC-Fr, ICC/IF
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-GFAP antibody [RM1003] - BSA and Azide free
    See all GFAP primary antibodies

  • Description

    Rabbit recombinant multiclonal [RM1003] to GFAP - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt (Intra), IHC-P, IP, IHC-Fr, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human, mouse and rat brain tissue lysates; U-87 MG, C6 whole cell lysates. IHC-P: Human, mouse and rat cerebrum tissue. IHC-Fr: Mouse cerebrum tissue. ICC/IF: Mouse primary neural/glia cell & rat hippocampal mixed glia. Flow Cyt (intra): Mouse primary brain cells. IP: Mouse brain lysate.

  • General notes

    ab283862 is the carrier-free version of ab278054.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab283862 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
WB
Use at an assay dependent concentration. Predicted molecular weight: 49 kDa.
Flow Cyt (Intra)
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
ICC/IF
Use at an assay dependent concentration.
Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 49 kDa.
Flow Cyt (Intra)
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
ICC/IF
Use at an assay dependent concentration.

Target

  • Function

    GFAP, a class-III intermediate filament, is a cell-specific marker that, during the development of the central nervous system, distinguishes astrocytes from other glial cells.

  • Tissue specificity

    Expressed in cells lacking fibronectin.

  • Involvement in disease

    Defects in GFAP are a cause of Alexander disease (ALEXD) [MIM:203450]. Alexander disease is a rare disorder of the central nervous system. It is a progressive leukoencephalopathy whose hallmark is the widespread accumulation of Rosenthal fibers which are cytoplasmic inclusions in astrocytes. The most common form affects infants and young children, and is characterized by progressive failure of central myelination, usually leading to death usually within the first decade. Infants with Alexander disease develop a leukoencephalopathy with macrocephaly, seizures, and psychomotor retardation. Patients with juvenile or adult forms typically experience ataxia, bulbar signs and spasticity, and a more slowly progressive course.

  • Sequence similarities

    Belongs to the intermediate filament family.

  • Post-translational
    modifications

    Phosphorylated by PKN1.

  • Cellular localization

    Cytoplasm. Associated with intermediate filaments.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • wu:fb34h11 antibody
    • ALXDRD antibody
    • cb345 antibody
    • etID36982.3 antibody
    • FLJ42474 antibody
    • FLJ45472 antibody
    • GFAP antibody
    • GFAP_HUMAN antibody
    • gfapl antibody
    • Glial fibrillary acidic protein antibody
    • Intermediate filament protein antibody
    • wu:fk42c12 antibody
    • xx:af506734 antibody
    • zgc:110485 antibody
    see all

Images

  • Western blot - Anti-GFAP antibody [RM1003] - BSA and Azide free (ab283862)
    Western blot - Anti-GFAP antibody [RM1003] - BSA and Azide free (ab283862)
    All lanes : Anti-GFAP antibody [RM1003] (ab278054) at 1/1000 dilution

    Lane 1 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate
    Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate
    Lane 3 : SK-N-BE (2) (Human neuroblastoma neuroblast) whole cell lysate
    Lane 4 : IMR-32 (Human lung fibroblast) whole cell lysate
    Lane 5 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 6 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate
    Lane 7 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
    Lane 8 : C6 (Rat glial tumor glial cell) whole cell lysate
    Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/20000 dilution

    Predicted band size: 49 kDa
    Observed band size: 40-54 kDa why is the actual band size different from the predicted?


    Exposure time: 180 seconds


    This data was developed using ab278054, the same antibody clone in a different buffer formulation. 

    Blocking and Diltuing buffer and concentration: 5% NFDM/TBST

    GFAP is expressed primarily by astrocytes in the CNS as astrocytes marker. We have no suitable astrocyte for western blot testing.
    U-87 MG and C6 were reported to express low level of GFAP (PMID: 23839947, PMID: 22096544, PMID: 20669222).

    NIH/3T3, IMR-32, HeLa and PC-12 were reported as negative cell lines for GFAP (PMID: 824020; PMID:2294; PMID: 6340792; PMID:9466565, PMID:7895062, PMID: 28700643, PMID: 19272755).

    No literature was found to support the expression in SH-SY5Y, Neuro-2a and SK-N-BE (2) cell.

     

  • Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    This data was developed using ab278054, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse cerebrum tissue labeling GFAP with ab278054 at 1/700 (0.938 µg/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at a 1/1000 dilution. Nuclear counterstain is DAPI. Positive staining on mouse cerebrum. 

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) used at a 1/1000 dilution.

    Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    This data was developed using ab278054, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling GFAP with ab278054 at 1/2000 (0.235 µg/ml) dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cerebrum. The section was incubated with ab278054 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [RM1003] - BSA and Azide free (ab283862)
    Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [RM1003] - BSA and Azide free (ab283862)
    This data was developed using ab278054, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cells labelling GFAP with ab278054 at 1/500 (0.938 µg/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green).
    Confocal image showing cytoplasmic staining in mouse primary astrocytes. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.

    ab10062 Anti-GFAP mouse monoclonal antibody at 1/200 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at a 1/500 dilution (Red) to counterstain. The nuclear counterstain was DAPI (Blue).

    Negative control 1: ab278054 at a 1/500 dilution followed by ab150120 at a 1/500 dilution.

    Negative control 2: ab10062 at a 1/500 dilution followed by ab150077 at a 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [RM1003] - BSA and Azide free (ab283862)
    Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [RM1003] - BSA and Azide free (ab283862)

    Immunofluorescence staining of GFAP using ab278054 in primary rat hippocampal mixed glia, (prepared from P2 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDPHP4m), DIV4. The cells were fixed with 100% MeOH (5 min), permeabilized with 0.1% Triton-X-100 (in PBS) for 5 mins 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 overnight at +4°C with ab278054 at 0.1 μg/ml and ab4674, Anti-GFAP antibody, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150176, Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown. The antibody ab278054 gave comparable results using 4% formaldehyde fixation (10 min).

    This data was developed using ab278054, the same antibody clone in a different buffer formulation.

  • Flow Cytometry (Intracellular) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    Flow Cytometry (Intracellular) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    This data was developed using ab278054, the same antibody clone in a different buffer formulation.

    Flow Cytometry analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized mouse primary brain cells labeling GFAP using ab278054 at a 1/500 dilution (0.1 µg) (Right panel) compared to Rabbit monoclonal IgG (ab172730) (Left panel).

    Secondary antibody is Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) used at a 1/2000 dilution.

  • Western blot - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    Western blot - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    All lanes : Anti-GFAP antibody [RM1003] (ab278054) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 49 kDa
    Observed band size: 40-54 kDa why is the actual band size different from the predicted?


    Exposure time: 3 seconds


    This data was developed using ab278054, the same antibody clone in a different buffer formulation. 

    The molecular weight observed is consistent with the literature (PMID: 25975510).

      

    Blocking/Dilution buffer: 5% NFDM/TBST.

     

  • Immunoprecipitation - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    Immunoprecipitation - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    This data was developed using ab278054, the same antibody clone in a different buffer formulation.

    GFAP was immunoprecipitated from 0.35 mg mouse brain lysate with ab278054 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab278054 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: Mouse brain lysate 10 μg.
    Lane 2: ab278054 IP in mouse brain lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab278054 in mouse brain lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 seconds.
    The molecular weight observed is consistent with the literature (PMID: 824020; PMID:2294; PMID: 6340792).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    This data was developed using ab278054, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling GFAP with ab278054 at 1/2000 (0.235 µg/ml) dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebrum. The section was incubated with ab278054 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Western blot - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    Western blot - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    All lanes : Anti-GFAP antibody [RM1003] (ab278054) at 1/1000 dilution

    Lane 1 : Human brain lysate
    Lane 2 : Human liver lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 49 kDa
    Observed band size: 40-54 kDa why is the actual band size different from the predicted?


    Exposure time: 3 seconds


    This data was developed using ab278054, the same antibody clone in a different buffer formulation. 

    Negative control: Human liver. (PMID: 25975510).

    The molecular weight observed is consistent with the literature (PMID: 25975510).

      

    Blocking/Diluting buffer: 5% NFDM/TBST.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    This data was developed using ab278054, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling GFAP with ab278054 at 1/2000 (0.235 µg/ml) dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat cerebrum. The section was incubated with ab278054 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)
    Anti-GFAP antibody [RP1003] - BSA and Azide free (ab283862)

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.

Click here to view the general protocols

References (0)

Publishing research using ab283862? Please let us know so that we can cite the reference in this datasheet.

ab283862 has not yet been referenced specifically in any publications.

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