Phosphothreonine | HUABIO HA723628 product information

product summary

company name :

HUABIO

product type :

antibody

product name :

Phosphothreonine

catalog :

HA723628

quantity :

100μl

price :

385.00 USD

clonality :

monoclonal

host :

domestic rabbit

conjugate :

nonconjugated

clone name :

PSH14-34

application :

western blot, ELISA, immunoprecipitation, immunohistochemistry - paraffin section

more info or order :

HUABIO product webpage

image

image 1 :

Western blot analysis of Phosphothreonine on different lysates with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/5,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate (20 µg/Lane) Observed band size: Mutiple 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 (HA723628) at 1/5,000 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.

image 2 :

Western blot analysis of Phosphothreonine on different lysates with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/5,000 dilution. Lane 1: NIH/3T3 cell lysate Lane 2: NIH/3T3 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate Lane 3: NIH/3T3 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour Lane 4: NIH/3T3 cell lysate Lane 5: NIH/3T3 treated with 1mM Sodium orthovanadate for 30 minutes cell lysate Lysates/proteins at 20 µg/Lane. Observed band size: Mutiple kDa Exposure time: 20 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 (HA723628) at 1/5,000 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.

image 3 :

Western blot analysis of Phosphothreonine on different lysates with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/5,000 dilution. Lane 1: PC-12 cell lysate Lane 2: PC-12 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate Lysates/proteins at 20 µg/Lane. Observed band size: Mutiple kDa Exposure time: 6 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 (HA723628) at 1/5,000 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.

product information

SKU :

HA723628

Target name :

Phosphothreonine

Species reactivity :

Human,Mouse,Rat,Species independent

Applications :

WB,IHC-P,IF-Cell,IP,ELISA

Conjugate :

Non-conjugated

Uniprot id :

Host :

Rabbit

Clone number :

PSH14-34

Isotype :

IgG

Size :

100μl

List Price :

385.00 USD

Storage Buffer :

PBS (pH7.4), 0.1% 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 :

HeLa treated with 100nM Calyculin A for 30 minutes cell lysate, NIH/3T3 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate, PC-12 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate, human gastric cancer tissue, human prostate tissue, human colon tissue, human liver tissue, human testis tissue, mouse prostate tissue, mouse colon tissue, mouse liver tissue, mouse testis tissue, rat colon tissue, rat liver tissue, rat testis tissue, HeLa, NIH/3T3, PC-12.

Concentration :

1 mg/mL.

Recommended dilutions :

WB: 1:5,000 ;IHC-P: 1:1,000-1:8,000 ;IF-Cell: 1:50,000 ;IP: 1-2μg/sample

Advanced Validation :

Cell treatment (CT)

Pic img4 :

https://storage.huabio.cn/huabio/productImg/HA723628_4.jpg

Pic legend4 :

Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue untreated / treated with λpp with Rabbit anti-Phosphothreonine antibody (HA723628) 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 (HA723628) 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 img5 :

https://storage.huabio.cn/huabio/productImg/HA723628_5.jpg

Pic legend5 :

Immunohistochemical analysis of paraffin-embedded human prostate tissue with Rabbit anti-Phosphothreonine antibody (HA723628) 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 (HA723628) 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 img6 :

https://storage.huabio.cn/huabio/productImg/HA723628_6.jpg

Pic legend6 :

Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/3,000 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 (HA723628) at 1/3,000 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 img7 :

https://storage.huabio.cn/huabio/productImg/HA723628_7.jpg

Pic legend7 :

Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/8,000 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 (HA723628) at 1/5,000 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 img8 :

https://storage.huabio.cn/huabio/productImg/HA723628_8.jpg

Pic legend8 :

Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/8,000 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 (HA723628) at 1/5,000 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 img9 :

https://storage.huabio.cn/huabio/productImg/HA723628_9.jpg

Pic legend9 :

Immunohistochemical analysis of paraffin-embedded mouse prostate tissue with Rabbit anti-Phosphothreonine antibody (HA723628) 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 (HA723628) 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 img10 :

https://storage.huabio.cn/huabio/productImg/HA723628_10.jpg

Pic legend10 :

Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/8,000 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 (HA723628) at 1/5,000 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 img11 :

https://storage.huabio.cn/huabio/productImg/HA723628_11.jpg

Pic legend11 :

Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/3,000 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 (HA723628) at 1/3,000 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/HA723628_12.jpg

Pic legend12 :

Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/3,000 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 (HA723628) at 1/3,000 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/HA723628_13.jpg

Pic legend13 :

Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/8,000 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 (HA723628) at 1/5,000 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 img14 :

https://storage.huabio.cn/huabio/productImg/HA723628_14.jpg

Pic legend14 :

Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/8,000 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 (HA723628) at 1/5,000 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 img15 :

https://storage.huabio.cn/huabio/productImg/HA723628_15.jpg

Pic legend15 :

Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/8,000 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 (HA723628) at 1/5,000 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 img16 :

https://storage.huabio.cn/huabio/productImg/HA723628_16.jpg

Pic legend16 :

Indirect ELISA analysis of Phosphothreonine.

Pic img17 :

https://storage.huabio.cn/huabio/productImg/HA723628_17.jpg

Pic legend17 :

Immunocytochemistry analysis of HeLa cells untreated / treated with 100nM Calyculin A for 30 minutes labeling Phosphothreonine with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/50,000 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-Phosphothreonine antibody (HA723628) at 1/50,000 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 (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.

Pic img18 :

https://storage.huabio.cn/huabio/productImg/HA723628_18.jpg

Pic legend18 :

Immunocytochemistry analysis of NIH/3T3 cells untreated / starved for 24 hours then treated with 100nM Calyculin A for 30 minutes labeling Phosphothreonine with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/50,000 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-Phosphothreonine antibody (HA723628) at 1/50,000 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 (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.

Pic img19 :

https://storage.huabio.cn/huabio/productImg/HA723628_19.jpg

Pic legend19 :

Immunocytochemistry analysis of PC-12 cells untreated / starved for 24 hours then treated with 100nM Calyculin A for 30 minutes labeling Phosphothreonine with Rabbit anti-Phosphothreonine antibody (HA723628) at 1/50,000 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-Phosphothreonine antibody (HA723628) at 1/50,000 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 (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.

Pic img20 :

https://storage.huabio.cn/huabio/productImg/HA723628_20.jpg

Pic legend20 :

Phosphothreonine was immunoprecipitated from 0.2 mg NIH/3T3 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate with HA723628 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723628 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: NIH/3T3 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate (input) Lane 2: HA723628 IP in NIH/3T3 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate Lane 3: Rabbit IgG instead of HA723628 in NIH/3T3 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 20 seconds; ECL: K1801

Pic img21 :

https://storage.huabio.cn/huabio/productImg/HA723628_21.jpg

Pic legend21 :

Phosphothreonine was immunoprecipitated from 0.2 mg PC-12 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate with HA723628 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723628 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: PC-12 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate (input) Lane 2: HA723628 IP in PC-12 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate Lane 3: Rabbit IgG instead of HA723628 in PC-12 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 25 seconds; ECL: K1801

more info or order :

HUABIO product webpage