Antibodies, Proteins, Kits and Reagents for Life Science

355 142 ₸ Product size

100 µl 355 142 ₸

Order now and get it on Tuesday March 02, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR1034Y] to GFAP
Suitable for: WB, IP, IHC-P, mIHC, ICC, IHC-Fr
Reacts with: Mouse, Rat, Human

Product name

Anti-GFAP antibody [EPR1034Y]
See all GFAP primary antibodies
Description

Rabbit monoclonal [EPR1034Y] to GFAP
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC	Mouse
IHC-Fr	Mouse Rat Human
IHC-P	Mouse Human
IP	Rat
mIHC	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide within Human GFAP aa 1 to the C-terminus (N terminal). The exact sequence is proprietary.
Positive control
- WB: Human, Mouse and Rat brain tissue lysate; Human, Mouse and Rat cerebullum tissue lysate; IHC-P: Mouse brain, cerebral cortex and liver tissue sections; Human brain, cerebral cortex, hippocampus, colon and glioma tissue sections; mIHC: Human cerebellum tissue; ICC: Mouse cerebellum and mouse primary neural mix culture cells; IP: Rat brain whole cell lysate. IHC-Fr: Human cerebral cortex tissue sections. Rat and Mouse cerebrum tissue sections.
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
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

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

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.21% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR1034Y
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

IHC image of GFAP staining in a formalin fixed, paraffin embedded normal human hippocampus tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab68428 at 1/100 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Immunocytochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural mix culture cells labelling GFAP with ab68428 at 1/250 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. ab10062 anti-GFAP antibody [GF5] at 1/100 dilution followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2µg/ml) was used as a counterstain. The Nuclear counterstain was DAPI (Blue).
Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebrum tissue labeling GFAP with ab68428 at 1/1000 dilution (2.011 µg/mL) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 µg/mL) (Green). Positive staining on rat cerebrum is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 µg/mL).
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)

All lanes : Anti-GFAP antibody [EPR1034Y] (ab68428) at 1/10000 dilution

Lane 1 : Human cerebellum tissue lysate at 20 µg
Lane 2 : Human brain tissue lysate at 10 µg

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 50 kDa
Observed band size: 48-50 kDa
why is the actual band size different from the predicted?

Blocking and Diluting buffer 5% NFDM/TBST
Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)

Fluorescence multiplex immunohistochemical analysis of human cerebellum tissue (formalin-fixed paraffin-embedded section).

Merged staining of Neu-N (ab177487; yellow; Opal™570), anti-beta III Tubulin (ab52623; red; Opal™690) and anti-GFAP (ab68428; green; Opal™520).

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ kit.

The section was incubated in three rounds of staining with ab177487 (1/1000 dilution), ab52623 (1/200 dilution) and ab68428 (1/250 dilution); each using a separate fluorescent tyramide signal amplification system.

Sodium citrate antigen retrieval (pH 6.0) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

DAPI (blue) was used as a nuclear counter stain.
Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum tissue labeling GFAP with ab68428 at 1/1000 dilution (2.011 µg/mL) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 µg/mL) (Green). Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 µg/mL).
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

IHC image of GFAP staining in a section of frozen normal human cerebral cortex performed on a Leica BOND^TM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab68428, 1/250 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)

All lanes : Anti-GFAP antibody [EPR1034Y] (ab68428) at 1/10000 dilution

Lane 1 : Mouse cerebellum tissue lysate
Lane 2 : Mouse brain tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 50 kDa
Observed band size: 50 kDa

Blocking and Diluting buffer 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue sections labelling GFAP with purified ab68428 at a dilution of 1/500. The secondary antibody used was ab97051, Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)

All lanes : Anti-GFAP antibody [EPR1034Y] (ab68428) at 1/50000 dilution

Lane 1 : Rat cerebellum tissue lysate
Lane 2 : Rat brain tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 50 kDa
Observed band size: 50 kDa

Blocking and Diluting buffer 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

Immunohistochemical analysis of paraffin-embedded human colon tissue sections labelling GFAP with purified ab68428 at a dilution of 1/500. The secondary antibody used was ab97051, Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0.
Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] (ab68428)

ab68428 at 1/20 dilution immunoprecipitating GFAP in rat brain whole cell lysate observed at 50 KDa (lanes 1 and 2).

Lane 1 (input): Rat brain whole cell lysate 10ug

Lane 2 (+): ab68428 + Rat brain whole cell lysate

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab68428 in Rat brain whole cell lysate

For western blotting, ab68428 was used followed by VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Blocking and Diluting buffer and concentration: 5% NFDM/TBST.
Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)

All lanes : Anti-GFAP antibody [EPR1034Y] (ab68428) at 1/5000 dilution (unpurified)

Lane 1 : Human brain lysate
Lane 2 : Rat brain lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled Goat anti-Rabbit antibody at 1/2000 dilution

Predicted band size: 50 kDa
Observed band size: 50 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue sections labelling GFAP with purified ab68428 at a dilution of 1/500. The secondary antibody used was ab97051, Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

Immunohistochemical analysis of formalin-fixed paraffin-embedded human brain (left) and human glioma (right) tissue sections labelling GFAP with unpurified ab68428 at dilution of 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue sections labelling GFAP with purified ab68428 at a dilution of 1/500. The secondary antibody used was ab97051, Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

Immunohistochemical analysis of formalin-fixed paraffin-embedded mouse brain tissue section labelling GFAP with unpurified ab68428 at dilution of 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Anti-GFAP antibody [EPR1034Y] (ab68428)

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