S100 alpha 6 Recombinant Rabbit Monoclonal Antibody [JF0976]
Catalog# ET1702-28
S100 alpha 6 Recombinant Rabbit Monoclonal Antibody [JF0976]
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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IP
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FC
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Human
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Mouse
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Rat
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HA750342
不含抗保成分
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unconjugated
Overview
Product Name
S100 alpha 6 Recombinant Rabbit Monoclonal Antibody [JF0976]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant full length protein of Human S100 alpha 6 aa 1-90 / 90.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Molecular Weight
Predicted band size: 10 kDa
Positive Control
HeLa cell lysate, A549 cell lysate, Neuro-2a cell lysate, C6 cell lysate, mouse lung tissue lysate, C6, human kidney tissue, human stomach carcinoma tissue, human colon carcinoma tissue, mouse brain tissue, mouse kidney tissue, rat kidney tissue.
Conjugation
unconjugated
Clone Number
JF0976
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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1:2,000-1:5,000
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IF-Cell
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1:100-1:500
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IF-Tissue
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1:100-1:500
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IHC-P
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1:50-1:1,000
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FC
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1:50-1:100
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IP
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Use at an assay dependent concentration.
Target
Function
Calcyclin, also known as Prolactin receptor-associated protein (PRA), growth factor-inducible protein 2A9, S-100 calcium-binding protein A6 (S-100A6) or MLN 4, is a homodimeric member of the S-100 calcium-binding protein family whose expression is upregulated in proliferating and differentiating cells. Calcyclin is inducible by growth factors and overexpressed in acute myeloid leukemias. It is expressed in a cell-specific manner in subpopulations of neurons and astrocytes and in epithelial cells and fibroblasts. Calcyclin is a specific target of S-100B protein in vivo. The binding of Calcyclin to S-100B is stabilized by S-100B-bound calcium and zinc. Calcyclin associates with both Annexin XI and CacyBP (calcyclin-binding protein). It functions to activate several processes along the calcium signal transduction pathway including the regulation of cell growth, proliferation, secretion and exocytosis.
Background References
1. Ralhan R et al. Immunohistochemical Subcellular Localization of Protein Biomarkers Distinguishes Benign from Malignant Thyroid Nodules: Potential for Fine-Needle Aspiration Biopsy Clinical Application. Thyroid 25:1224-34 (2015).
2. Alves RM et al. iTRAQ-based quantitative proteomic analysis of submandibular glands from rats with STZ-induced hyperglycemia. J Biochem 153:209-20 (2013).
Sequence Similarity
Belongs to the S-100 family.
Post-translational Modification
The N-terminus is blocked.
Subcellular Location
Nucleus envelope, Cytoplasm, Cell membrane.
Synonyms
2A9 antibody
5B10 antibody
CABP antibody
CACY antibody
Calcyclin antibody
Growth factor inducible protein 2A9 antibody
Growth factor-inducible protein 2A9 antibody
MLN 4 antibody
MLN4 antibody
OTTHUMP00000015472 antibody
ExpandImages
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Western blot analysis of S100 alpha 6 on different lysates with Rabbit anti-S100 alpha 6 antibody (ET1702-28) at 1/2,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: A549 cell lysate
Lane 3: Neuro-2a cell lysate
Lane 4: C6 cell lysate
Lane 5: Mouse lung tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 10 kDa
Observed band size: 10 kDa
Exposure time: 10 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 (ET1702-28) at 1/2,000 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. -
☑ Knockdown (KD)
Western blot analysis of S100 alpha 6 on different lysates with Rabbit anti-S100 alpha 6 antibody (ET1702-28) at 1/2,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-S100 alpha 6 KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 10 kDa
Observed band size: 10 kDa
Exposure time: 1 minute; 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 (ET1702-28) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of C6 cells labeling S100 alpha 6 with Rabbit anti-S100 alpha 6 antibody (ET1702-28) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-S100 alpha 6 antibody (ET1702-28) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-S100 alpha 6 antibody (ET1702-28) at 1/1,000 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 (ET1702-28) at 1/1,000 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 human kidney tissue using anti-S100 alpha 6 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1702-28, 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 stomach carcinoma tissue using anti-S100 alpha 6 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1702-28, 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 colon carcinoma tissue using anti-S100 alpha 6 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1702-28, 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 mouse brain tissue using anti-S100 alpha 6 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1702-28, 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 mouse kidney tissue using anti-S100 alpha 6 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1702-28, 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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Flow cytometric analysis of S100 alpha 6 was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1702-28, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.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"