Cyclin D1 | HUABIO ET1601-31 product information

product summary

company name :

HUABIO

product type :

antibody

product name :

Cyclin D1

catalog :

ET1601-31

quantity :

100μl

price :

385.00 USD

clonality :

monoclonal

host :

domestic rabbit

conjugate :

nonconjugated

clone name :

SA38-08

reactivity :

human, mouse, rat

application :

western blot, immunoprecipitation, flow cytometry, immunohistochemistry - paraffin section

more info or order :

HUABIO product webpage

image

image 1 :

Western blot analysis of Cyclin D1 on different lysates with Rabbit anti-Cyclin D1 antibody (ET1601-31) at 1/5,000 dilution and competitor's antibody at 1/5,000 dilution. Lane 1: MCF7 cell lysate Lane 2: K-562 cell lysate (negative) Lane 3: A431 cell lysate Lane 4: Neuro-2a cell lysate Lane 5: NIH/3T3 cell lysate Lane 6: C6 cell lysate Lane 7: SH-SY5Y cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 34 kDa Observed band size: 35 kDa Exposure time: 20 seconds; ECL: K1802; 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 (ET1601-31) at 1/5,000 dilution and competitor's antibody at 1/5,000 dilution were 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.

image 2 :

Western blot analysis of Cyclin D1 on different lysates with Rabbit anti-Cyclin D1 antibody (ET1601-31) at 1/5,000 dilution. Lane 1: MCF7-si NT cell lysate Lane 2: MCF7-si Cyclin D1 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 34 kDa Observed band size: 34 kDa Exposure time: 17 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 (ET1601-31) at 1/5,000 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.

image 3 :

Cyclin D1 was immunoprecipitated from 0.5 mg Hela whole cell lysates with ET1601-31 at 2 μg/mL. Western blot was performed from the immunoprecipitate using ET1601-31 at 1/500 dilution for 45 minutes at room temperature. Goat anti-Rabbit IgG-HRP Secondary Antibody (HA1001) was used at 1:300,000 dilution for 30 minutes at room temperature. Lane 1: Hela whole cell lysates at 10 μg; Lane 2: Cyclin D1 (ET1601-31) IP in Hela whole cell lysates; Lane 3: Rabbit IgG instead of Cyclin D1 (ET1601-31) in Hela whole cell lysates. Predicted band size: 34 kDa Observed band size: 34 kDa Exposure time: 5 minutes; 12% SDS-PAGE gel.

product information

SKU :

ET1601-31

Target name :

Cyclin D1

Species reactivity :

Human,Mouse,Rat

Applications :

WB,IF-Cell,IF-Tissue,IHC-P,IP,FC

Conjugate :

Non-conjugated

Immunogen :

Synthetic peptide within C-terminal human Cyclin D1.

Uniprot id :

P24385>SwissProt: P24385 Human;SwissProt: P25322 Mouse;SwissProt: P39948 Rat

Host :

Rabbit

Clone number :

SA38-08

Isotype :

IgG

Size :

100μl

List Price :

385.00 USD

Storage Buffer :

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Form :

Liquid

Storage Instruction :

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Purity :

Protein A affinity purified.

Product type :

Recombinant Rabbit monoclonal Antibody

Positive control :

MCF7 cell lysate, K-562 cell lysate, A431 cell lysate, Neuro-2a cell lysate, NIH/3T3 cell lysate, C6 cell lysate, SH-SY5Y cell lysate, Neuro-2a, MCF7, human tonsil tissue, human colon carcinoma tissue, human liver carcinoma tissue, human small intestine tissue.

Molecular wt :

Predicted band size: 34 kDa

Subcellular location :

Cytoplasm, Nucleus, Membrane, Mitochondrion

Concentration :

1 mg/mL.

Recommended dilutions :

WB: 1:5,000 ;IF-Cell: 1:2,000 ;IF-Tissue: 1:500 ;IHC-P: 1:200-1:1,000 ;IP: Use at an assay dependent concentration. ;FC: 1:5,000

Advanced Validation :

Relative expression (RE),Knockdown (KD)

Pic img4 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_4.jpg

Pic legend4 :

Immunocytochemistry analysis of Neuro-2a cells labeling Cyclin D1 with Rabbit anti-Cyclin D1 antibody (ET1601-31) at 1/2,000 dilution and competitor's antibody at 1/1,600 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Cyclin D1 antibody (ET1601-31) at 1/2,000 dilution and competitor's antibody at 1/1,600 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 (M1305-2, 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.

Pic img5 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_5.jpg

Pic legend5 :

Flow cytometric analysis of MCF7 cells labeling Cyclin D1. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-31, red) at 1/5,000 dilution and competitor's antibody (red) at 1/2,000 dilution, 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).

Pic img6 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_6.jpg

Pic legend6 :

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Cyclin D1 antibody (ET1601-31) at 1/1,000 dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. The section was incubated with ET1601-31 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img7 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_7.jpg

Pic legend7 :

Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Cyclin D1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1601-31, 1/200) 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.

Pic img8 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_8.jpg

Pic legend8 :

Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Cyclin D1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1601-31, 1/200) 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.

Pic img9 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_9.jpg

Pic legend9 :

Immunohistochemical analysis of paraffin-embedded human small intestine tissue using anti-Cyclin D1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1601-31, 1/200) 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.

Pic img10 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_10.jpg

Pic legend10 :

Pic img11 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_11.jpg

Pic legend11 :

Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-Cyclin D1 antibody (ET1601-31) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1601-31) 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.

Pic img12 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_12.jpg

Pic legend12 :

Immunohistochemical analysis of paraffin-embedded rat esophagus tissue with Rabbit anti-Cyclin D1 antibody (ET1601-31) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1601-31) 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.

Pic img13 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_13.jpg

Pic legend13 :

Application: IF-Tissue Species: Human Site: colon cancer Sample: Paraffin-embedded section Antibody concentration: 1/500

Pic img14 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_14.jpg

Pic legend14 :

Immunocytochemistry analysis of SH-SY5Y (positive) and 293T (negative) labeling Cyclin D1 with Rabbit anti-Cyclin D1 antibody (ET1601-31) at 1/2,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Cyclin D1 antibody (ET1601-31) at 1/2,000 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 (M1305-2, 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.

Pic img15 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_15.jpg

Pic legend15 :

Immunocytochemistry analysis of C6 cells labeling Cyclin D1 with Rabbit anti-Cyclin D1 antibody (ET1601-31) at 1/2,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Cyclin D1 antibody (ET1601-31) at 1/2,000 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 (M1305-2, 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.

Pic img16 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_16.jpg

Pic legend16 :

Cyclin D1 was immunoprecipitated from 0.2 mg MCF7 cell lysate with ET1601-31 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1601-31 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: MCF7 cell lysate (input) Lane 2: ET1601-31 IP in MCF7 cell lysate Lane 3: Rabbit IgG instead of ET1601-31 in MCF7 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 5 seconds; ECL: K1802

Pic img17 :

https://storage.huabio.cn/huabio/productImg/ET1601-31_17.jpg

Pic legend17 :

Immunohistochemical analysis of paraffin embedded human colon cancer tissue using anti-Cyclin D1 antibody (1/1,000) performed on the Leica® BOND™ RX.

more info or order :

HUABIO product webpage