Antibodies, Proteins, Kits and Reagents for Life Science

288 134 ₸ Product size

100 µl 288 134 ₸

Order now and get it on Tuesday March 02, 2021

Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR22035-112] to 68kDa Neurofilament/NF-L
Suitable for: WB, IHC-P, IHC-Fr, IP, ICC/IF
Reacts with: Mouse, Rat, Human

Product name

Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112]
See all 68kDa Neurofilament/NF-L primary antibodies
Description

Rabbit monoclonal [EPR22035-112] to 68kDa Neurofilament/NF-L
Host species

Rabbit

Tested Applications & Species

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

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human, mouse and rat brain lysate. IHC-P: Human, mouse and rat cerebellum tissue. IHC-Fr: Mouse and rat cerebellum tissue. IP: Mouse brain lysate. ICC/IF Mouse primary neuron cells
General notes
Previously labelled as 68kDa Neurofilament.
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.
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 long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22035-112
Isotype

IgG
Research areas

Immunohistochemistry (Frozen sections) - Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen sectioned mouse cerebellum tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution (green). Positive staining in mouse cerebellum (PMID: 17598884) is observed. The nuclear counter stain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20).
Immunocytochemistry/ Immunofluorescence - Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343)

Immunocytochemistry/ Immunofluorescence analysis of mouse primary neuron cells labeling Neurofilament/NF-L with purified ab223343 at 1/50 (9.4µg/mL). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab11267 Anti-MAP2 antibody [HM-2]; ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.
Western blot - Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343)

All lanes : Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343) at 1/1000 dilution

Lane 1 : Human brain lysate
Lane 2 : Mouse brain lysate
Lane 3 : Rat brain lysate
Lane 4 : Mouse 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: 62 kDa
Observed band size: 68 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking and dilution buffer: 5% NFDM/TBST.

Negative control: Mouse liver (PMID: 2120242).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343)

Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining of nerve fibers in rat cerebellum (PMID: 28190929) is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Frozen sections) - Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen sectioned rat cerebellum tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution (green). Positive staining in rat cerebellum is observed. The nuclear counter stain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 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-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343)

Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining of nerve fibers in mouse cerebellum (PMID: 28190929) is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343)

Immunohistochemical analysis of paraffin-embedded human cerebellum tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining of nerve fibers in human cerebellum (PMID: 28190929) is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunoprecipitation - Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343)

68kDa Neurofilament/NF-L was immunoprecipitated from 0.35 mg of mouse brain lysate with ab223343 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab223343 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1: Mouse brain lysate 10 μg (Input).
Lane 2: ab223343 IP in mouse brain lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab223343 in mouse brain lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
Immunohistochemistry (Frozen sections) - Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen sectioned rat liver tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution. No staining in rat liver is observed. The nuclear counter stain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

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

Negative control :Rat liver (PMID: 2120242).
Immunohistochemistry (Frozen sections) - Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen sectioned mouse liver tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution. No staining observed in mouse liver. The nuclear counter stain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

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

Negative control: mouse liver (PMID: 2120242).
Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343)

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