Alpha B Crystallin Recombinant Rabbit Monoclonal Antibody [JA50-32]
Catalog# ET1704-60
Alpha B Crystallin Recombinant Rabbit Monoclonal Antibody [JA50-32]
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WB
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IHC-P
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IF-Cell
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IF-Tissue
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IP
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FC
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Human
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Rat
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unconjugated
Overview
Product Name
Alpha B Crystallin Recombinant Rabbit Monoclonal Antibody [JA50-32]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human Alpha B Crystallin aa 126-175 / 175.
Species Reactivity
Human, Rat
Validated Applications
WB, IHC-P, IF-Cell, IF-Tissue, IP, FC
Molecular Weight
Predicted band size: 20 kDa
Positive Control
U-2 OS cell lysate, rat heart tissue lysate, human skeletal muscle tissue lysate, Hela, human embryonic skeletal muscle tissue, human prostate tissue, human cardiac muscle tissue, rat cardiac muscle tissue, U-2 OS.
Conjugation
unconjugated
Clone Number
JA50-32
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:500-1:2,000
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IF-Cell
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1:50-1:200
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IF-Tissue
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1:50-1:200
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IHC-P
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1:50-1:200
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IP
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Use at an assay dependent concentration.
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FC
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1:1,000
Target
Function
Crystallins are the major proteins of the vertebrate eye lens, where they maintain the transparency and refractive index of the lens. Crystallins are divided into α, β and γ families, and the β- and γ-crystallins also compose a superfamily. Crystallins usually contain seven distinct protein regions, inclu-ding four homologous motifs, a connecting peptide, and N- and C-terminal extensions. α-crystallins consist of three gene products, αA-, αB- and αC-crystallin, which are members of the small heat shock protein family (HSP 20). α-crystallins act as molecular chaperones by holding denatured proteins in large soluble aggregates. However, unlike other molecular chaperones, α-crystallins do not renature these proteins. Expression of αA-crystallin is restricted to the lens and defects of this gene cause the development of autosomal dominant congenital cataracts (ADCC). The human αB-crystallin gene product is expressed in many tissues, including lens, heart and skeletal muscle. Elevated expression of αB-crystallin is associated with many neurological diseases, and a missense mutation in this gene has co-segregated in a family with a Desmin-related myopathy.
Background References
1. Sosunov, A.A. et al. Phenotypic conversions of "protoplasmic" to "reactive" astrocytes in Alexander disease. The Journal of neuroscience : the official journal of the Society for Neuroscience. 33: 7439-50 (2013).
2. Lambrecht S et al. Differential expression of alphaB-crystallin and evidence of its role as a mediator of matrix gene expression in osteoarthritis. Arthritis Rheum 60:179-88 (2009).
Sequence Similarity
Belongs to the small heat shock protein (HSP20) family.
Tissue Specificity
Lens as well as other tissues. Expressed in myocardial tissue.
Subcellular Location
Nucleus, Cytoplasm.
Synonyms
AACRYA antibody
Alpha B crystallin antibody
Alpha crystallin B chain antibody
Alpha(B)-crystallin antibody
Alpha-crystallin B chain antibody
CRYA2 antibody
Cryab antibody
CRYAB_HUMAN antibody
Crystallin alpha B antibody
Crystallin alpha polypeptide 2 antibody
ExpandImages
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Western blot analysis of Alpha B Crystallin on different lysates with Rabbit anti-Alpha B Crystallin antibody (ET1704-60) at 1/1,000 dilution.
Lane 1: U-2 OS cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 20 kDa
Observed band size: 20 kDa
Exposure time: 59 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-60) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of Alpha B Crystallin on different lysates with Rabbit anti-Alpha B Crystallin antibody (ET1704-60) at 1/1,000 dilution.
Lane 1: Rat heart tissue lysate
Lysates/proteins at 40 µg/Lane.
Predicted band size: 20 kDa
Observed band size: 20 kDa
Exposure time: 2 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-60) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of Alpha B Crystallin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1704-60, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:20,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Rat heart tissue lysate
Lane 2: Human skeletal muscle tissue lysate -
ICC staining of Alpha B Crystallin in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-60, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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Immunohistochemical analysis of paraffin-embedded human embryonic skeletal muscle tissue using anti-Alpha B Crystallin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-60, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human prostate tissue using anti-Alpha B Crystallin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-60, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human cardiac muscle tissue with Rabbit anti-Alpha B Crystallin antibody (ET1704-60) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-60) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat cardiac muscle tissue with Rabbit anti-Alpha B Crystallin antibody (ET1704-60) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-60) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of MCF7 (left, negative) and U-2 OS (right, positive) cells labeling Alpha B Crystallin.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1704-60, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"