Purified anti-Alpha-II Spectrin Antibody

Pricing & Availability
Clone
D8B7 (See other available formats)
Other Names
Spectrin alpha chain, brain, alpha-II spectrin, fodrin alpha chain, spectrin, non-erythroid alpha chain, EL2, SPTA1
Isotype
Mouse IgG2b, κ
Ave. Rating
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Product Citations
publications
D8B7_Pure_AlphaIISpectrin_Antibody_1_IHC_073118
IHC staining of purified anti-Alpha-II Spectrin antibody (clone D8B7) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R (Cat. No. 928602), the tissue was incubated with 1 µg/ml of the primary antibody overnight at 4°C. BioLegend´s Ultra-Streptavidin (USA) HRP kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective. Scale bar: 50 µm
  • D8B7_Pure_AlphaIISpectrin_Antibody_1_IHC_073118
    IHC staining of purified anti-Alpha-II Spectrin antibody (clone D8B7) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R (Cat. No. 928602), the tissue was incubated with 1 µg/ml of the primary antibody overnight at 4°C. BioLegend´s Ultra-Streptavidin (USA) HRP kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective. Scale bar: 50 µm
  • D8B7_Pure_AlphaIISpectrin_Antibody_2_IHC_073118
    IHC staining of purified anti-Alpha-II Spectrin antibody (clone D8B7) on formalin-fixed paraffin-embedded rat brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R (Cat. No. 928602), the tissue was incubated with 1 µg/ml of the primary antibody overnight at 4°C. BioLegend´s Ultra-Streptavidin (USA) HRP kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective. Scale bar: 50 µm
  • D8B7_Pure_AlphaIISpectrin_Antibody_3_ICC_073118
    ICC staining of purified anti-Alpha-II Spectrin antibody (clone D8B7) on HeLa cells. The cells were fixed with 4% PFA, permeabilized with a buffer containing 0.1% Triton X-100 and 0.25% BSA, and blocked with 2% normal goat serum and 0.02% BSA. The cells were then incubated with 1 µg/ml of the primary antibody for overnight at 4°C, followed by incubation with 2.5 µg/ml of Alexa Fluor® 594 goat anti-Mouse IgG for one hour at room temperature. Nuclei were counterstained with DAPI, and the slides were mounted with ProLong™ Gold Antifade Mountant. The image was captured with a 40X objective. Scale bar: 50 µm
  • D8B7_Pure_AlphaIISpectrin_Antibody_4_ICC_073118
    ICC staining of purified anti-Alpha-II Spectrin antibody (clone D8B7) on 3T3 cells. The cells were fixed with 4% PFA, permeabilized with a buffer containing 0.1% Triton X-100 and 0.25% BSA, and blocked with 2% normal goat serum and 0.02% BSA. The cells were then incubated with 1 µg/ml of the primary antibody for overnight at 4°C, followed by incubation with 2.5 µg/ml of Alexa Fluor® 594 goat anti-Mouse IgG for one hour at room temperature. Nuclei were counterstained with DAPI, and the slides were mounted with ProLong™ Gold Antifade Mountant. The image was captured with a 40X objective. Scale bar: 50 µm
  • D8B7_Pure_AlphaIISpectrin_Antibody_5_WB_073118
    Western blot of purified anti-Alpha-II Spectrin antibody (clone D8B7). Lane 1: Molecular weight marker; Lane 2: 20 µg of human brain lysate; Lane 3: 20 µg of mouse brain lysate; Lane 4: 20 µg of rat brain lysate; Lane 5: 20 µg of 3T3 cell lysate; Lane 6: 20 µg of HeLa cell lysate. The blot was incubated with 0.5 µg/mL of the primary antibody overnight at 4°C, followed by incubation with HRP labeled goat anti-mouse IgG (Cat. No. 405306). Enhanced chemiluminescence was used as the detection system.
  • D8B7_Pure_AlphaIISpectrin_Antibody_6_WB_051519
    Western blot of purified anti-Alpha-II Spectrin antibody (clone D8B7). Lane 1: Molecular weight marker; Lane 2: 20 µg of Drosophila head lysate; Lane 3: 20 µg of Drosophila S2 (embryonic) cell lysate. The blot was incubated with 0.1 µg/mL of the primary antibody overnight at 4°C, followed by incubation with HRP-labeled goat anti-mouse IgG (Cat. No. 436302). Enhanced chemiluminescence was used as the detection system.
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803205 25 µg $95
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803206 100 µg $245
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Description

The spectrins are a family of widely distributed cytoskeletal proteins which are involved in actin crosslinking. Spectrin α-chain heterodimerizes with tissue-specific β-chains. It is specifically expressed in nonerythrocytic cells. The protein has been implicated in other cellular functions including DNA repair and cell cycle regulation. Mutations in the gene are the cause of early infantile epileptic encephalopathy. Fodrin, a spectrin that seems to be involved in secretion, interacts with calmodulin in a calcium-dependent manner and is thus a candidate for the calcium-dependent movement of the cytoskeleton at the membrane.

Product Details
Technical data sheet

Product Details

Reactivity
Human, Mouse, Rat, Drosophila
Antibody Type
Monoclonal
Host Species
Mouse
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

IHC-P - Quality tested
ICC, WB - Validated

Recommended Usage

Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 1.0 - 5.0 µg/ml is suggested. For immunocytochemistry, a concentration of 1.0 μg/ml is recommended. For Western blotting, the suggested use of this reagent is 0.05 - 2.0 µg per ml. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Clone D8B7 is a specific marker for human, rat and mouse alpha II spectrin or fodrin. In cerebellar cell cultures, it labels axons and periphery of cell bodies consistent with localization of fodrin (reviewed in Bennett, 1993; Goodman, 1995 & De Matteis, 2000)4, 6, 8. It does not cross-react with alpha I (erythroid) spectrin SH3 domain.

This antibody was developed against a recombinant alpha II spectrin (non-erythroid spectrin or fodrin) SH3 domain. It was subsequently purified via ammonium sulfate precipitation.

Reactivity to Drosophila was only validated with the purified format.

Additional Product Notes

For Western blotting, the suggested use of this reagent is 0.05 - 1 µg/ml in human, mouse, and rat and 0.1 - 2 µg/ml in Drosophila.

Application References

(PubMed link indicates BioLegend citation)
  1. Konieczny P, et al. 2008. J Cell Biol. 181:667-81
  2. Xu J, et al. 2001. Brain Res. 898:171-7 (WB, IF)
  3. Xu J, et al. 2000. J Cell Science. 113:3805-14
  4. De Matteis MA and Morrow JS. 2000. J. Cell Science. 113:2331-43
  5. Swanson JA and Watts C. 1995. Trends Cell Biol. 5:424-427
  6. Goodman SR, et al. 1995. Brain Res Bull. 36:593-606
  7. Lamaze C and Schimid SL. 1995. Curr Opin Cell Biol. 7:573-580
  8. Bennett V and Gilligan DM. 1993. Annu Rev Cell Biol. 9:27-66
  9. Huang C, et al. 2017. J Neurosci. 37:11323-11334 PubMed
RRID
AB_2749952 (BioLegend Cat. No. 803205)
AB_2749953 (BioLegend Cat. No. 803206)

Antigen Details

Structure
Alpha-II Spectrin is a 2472 amino acid protein with an expected molecular mass of 285 kD. A cleavage product of ~ 150 kD is observed in tissue lysates (1) (2).
Biology Area
Cell Biology, Cell Motility/Cytoskeleton/Structure, Neuroscience, Neuroscience Cell Markers
Molecular Family
Presynaptic proteins, Scaffold Proteins
Antigen References
  1. Pike BR, et al. 2001. J Neurochem. 78(6):1297-306.
  2. Weiss ES, et al. 2009. Ann Thorac Surg. 88(2):543
Gene ID
6709 View all products for this Gene ID
UniProt
View information about Alpha-II Spectrin on UniProt.org

Related FAQs

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Go To Top Version: 2    Revision Date: 05/15/2019

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