JNK1+2+3 (T183+Y185) Polyclonal Antibody

$L1 mouse brain lysates L2 human colon carcinoma lysates probed with Anti - phospho-JNK1\/2\/3 (Thr183+Tyr185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 in 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in 37˚C. Predicted band 42kD. Observed band size: 42kD$

L1 mouse brain lysates L2 human colon carcinoma lysates probed with Anti - phospho-JNK1\/2\/3 (Thr183+Tyr185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 in 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in 37˚C. Predicted band 42kD. Observed band size: 42kD

Image kindly provided by Dr. Magdalena Krol. Control tumor cells, tumor cells grown in macrophage-conditioned medium, tumor cells sorted from co-culture with macrophages, and macrophages from monocultures and sorted from co-culture with tumor cells were analyzed. Total protein concentrations in lysates were determined using a Bio-Rad protein assay. Proteins (50 mg) were resolved using SDS-PAGE and transferred onto PVDF membranes. The membranes were then blocked with 5% non-fat dry milk in TBS buffer containing 0.5% Tween 20. The membranes were then incubated overnight with the primary Rabbit Anti-JNK1+2+3 (THR183+TYR185) at 1:100 dilution. Subsequently, the membranes were washed three times in TBS containing 0.5% Tween 20 and incubated for 1 h at room temperature with secondary antibodies conjugated with the appropriate infrared (IR) fluorophore IRDyeH 800 CW or IRDyeH 680 RD at a dilution of 1:5000.

$Mouse splenocytes probed with Rabbit Anti-JNK1\/2\/3 (Thr183+Tyr185) Polyclonal Antibody, Unconjugated (bs-1640R) (green) at 1:50 for 30 minutes followed by a FITC conjugated secondary antibody compared to unstained cells (blue), secondary only(light blue), and isotype control(orange).$

Mouse splenocytes probed with Rabbit Anti-JNK1\/2\/3 (Thr183+Tyr185) Polyclonal Antibody, Unconjugated (bs-1640R) (green) at 1:50 for 30 minutes followed by a FITC conjugated secondary antibody compared to unstained cells (blue), secondary only(light blue), and isotype control(orange).

$Paraformaldehyde-fixed, paraffin embedded Human brain glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:400 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Human brain glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:400 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:400 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:400 overnight at 4\u00b0C, DAB staining.

Lane 1: Rat Cerebrum lysates; Lane 2: Mouse Cerebrum lysates; Lane 3: Rat Cerebellum lysates; Lane 4: Mouse Cerebellum lysates; Lane 5: Rat Heart lysates; Lane 6: Mouse Heart lysates; Lane 7: Mouse Kidney lysates probed with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

$Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Mouse cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Mouse heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Rat kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Rat cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Human colon cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Human colon cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.

Lane 1: Rat Cerebrum lysates; Lane 2: Mouse Cerebrum lysates; Lane 3: Rat Cerebellum lysates; Lane 4: Mouse Cerebellum lysates; Lane 5: Rat Heart lysates; Lane 6: Mouse Heart lysates; Lane 7: Mouse Kidney lysates probed with JNK1+2+3 Polyclonal Antibody, Unconjugated (bs-1640R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

$Jurkat cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with JNK1+2+3 (T183+Y185) Antibody(bs-1640R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

Jurkat cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with JNK1+2+3 (T183+Y185) Antibody(bs-1640R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

$Paraformaldehyde-fixed, paraffin embedded Human brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Human brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Human gastric carcinoma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Human gastric carcinoma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Mouse kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with JNK1+2+3 (T183+Y185) Polyclonal Antibody, Unconjugated (bs-1640R) at 1:200 overnight at 4\u00b0C, DAB staining.

Applications

WB
ELISA
FCM
IHC-P
IHC-F
IF(IHC-P)
IF(IHC-F)
IF(ICC)
ICC

Reactivity

Human
Mouse
Rat
Dog
Rabbit

Predicted Reactivity

Overview
Catalog #	bs-1640R
Product Name	JNK1+2+3 (T183+Y185) Polyclonal Antibody
Applications	WB, ELISA, FCM, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC), ICC
Reactivity	Human, Mouse, Rat, Dog, Rabbit
Predicted Reactivity	Cow, Pig
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic phosphopeptide derived from human JNK1 around the phosphorylation site of Thr183/Tyr185
Modification Site	THR183+TYR185
Clonality	Polyclonal
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	5599
Swiss Prot	P45983
Subcellular location	Cytoplasm, Nucleus
Synonyms	JNK; JNK1; PRKM8; SAPK1; JNK-46; JNK1A2; SAPK1c; JNK21B1/2; Mitogen-activated protein kinase 8; MAP kinase 8; MAPK 8; Stress-activated protein kinase 1c; Stress-activated protein kinase JNK1; c-Jun N-terminal kinase 1; MAPK8
Background	JNK1 isoforms display different binding patterns: beta-1 preferentially binds to c-Jun, whereas alpha-1, alpha-2, and beta-2 have a similar low level of binding to both c-Jun or ATF2. However, there is no correlation between binding and phosphorylation, which is achieved at about the same efficiency by all isoforms.
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
FCM	1:20-100
IHC-P	1:200-400
IHC-F	1:100-500
IF(IHC-P)	1:50-200
IF(IHC-F)	1:50-200
IF(ICC)	1:50-200
ICC	1:100-500