710928

antibody from Invitrogen Antibodies

Targeting: JAK2 JTK10

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

710928

Provider

Invitrogen Antibodies

Product name

Phospho-JAK2 (Tyr1007, Tyr1008) Recombinant Polyclonal Antibody (18HCLC)

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Monkey, Pig, Sheep and Bovine.

Antibody clone number

18HCLC

Concentration

0.5 mg/mL

References

Submitted references

Xanthotoxin and umbelliferone attenuate cognitive dysfunction in a streptozotocin-induced rat model of sporadic Alzheimer's disease: The role of JAK2/STAT3 and Nrf2/HO-1 signalling pathway modulation.

Hindam MO, Sayed RH, Skalicka-Woźniak K, Budzyńska B, El Sayed NS
Phytotherapy research : PTR 2020 Sep;34(9):2351-2365

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ATM‑JAK‑PD‑L1 signaling pathway inhibition decreases EMT and metastasis of androgen‑independent prostate cancer.

Zhang L, Xu LJ, Zhu J, Li J, Xue BX, Gao J, Sun CY, Zang YC, Zhou YB, Yang DR, Shan YX
Molecular medicine reports 2018 May;17(5):7045-7054

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of HEL 92.1.7 (1) and HEL 92.1.7 treated with MG132 (20 uM, 1 hour) followed by IFN-gamma (100 ng, 15 min) (2). The blots were probed with Anti-Jak (pY1007/1008) Recombinant Rabbit Polyclonal Antibody (Product # 710928, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 131 kDa band corresponding to Jak (pY1007/1008) was observed. To confirm the specificity of Anti-Jak (pY1007/1008) Recombinant Rabbit Polyclonal Antibody, competition was performed with the non-phosphopeptide (10 µg/mL) and phosphopeptide (10 µg/mL) as shown in the corresponding blot on the right and extrme right respectively. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with overnight wet transfer system. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis HT-29 cells were fixed and permeabilized for detection of Jak (pY1007/1008) using Jak (pY1007/1008) Recombinant Rabbit Polyclonal Antibody (Product # 710928, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938), and Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Detection and localization of Jak (pY1007/1008) (green) in the cytoplasmic can be clearly observed in cells treated with MG132 (20 uM, 1 hr) and IFN-gamma (100 ng, 15 min) as compared to untreated cells. Antibody specificity was demonstrated by competition with the Jak (pY1007/1008) peptide, which results in loss of signal. No competition was observed with the non-phospho peptide.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4. Levels of PD-L1, p-JAK1, p-JAK2 and p-STAT3 are suppressed in C4-2-ATMSi and CWR22Rv1-ATMSi cells compared with in the control cells, and JAK inhibitor 1 significantly suppresses the expression of PD-L1 in ATM knockout groups and control groups. (A) A significant decrease in PD-L1 expression was revealed in experimental groups by reverse transcription-quantitative polymerase chain reaction and western blotting. (B) Decreased levels of p-JAK1, p-JAK2 and p-STAT3 were revealed in the experimental groups by western blotting. (C) JAK inhibitor 1 and Stattic were used to treat all cell groups. Downregulation of JAK significantly reduced PD-L1 expression, whereas Stattic had no significant effect on PD-L1 expression. **P