Anti-Kallikrein 1/KLK1 Antibody Picoband®

Anti-Kallikrein 1 antibody, PA1709, Western blotting
Lane 1: Recombinant Mouse KLK1 Protein 10ng
Lane 2: Recombinant Mouse KLK1 Protein 5ng
Lane 3: Recombinant Mouse KLK1 Protein 2.5ng
Lane 4: Recombinant Mouse KLK1 Protein 1.25ng

Western blot analysis of KLK1 using anti-KLK1 antibody (PA1709).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat pancreas tissue lysates,
Lane 2: mouse pancreas tissue lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-KLK1 antigen affinity purified polyclonal antibody (Catalog # PA1709) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for KLK1 at approximately 29KD. The expected band size for KLK1 is at 29KD.

Anti-Kallikrein 1 antibody, PA1709, IHC(P)
IHC(P): Mouse Kidney Tissue

Acinar cell apoptosis is delayed in post-ligated SMGs of ATG5-deficient mice. ( a ) ATG5 status impinges upon duct ligation-triggered acinar apoptosis. Apoptosis by ApopTag Peroxidase (brown nuclear staining) was visualized using an In situ Apoptosis Detection Kit ( left panel ). Peak apoptosis was detected in L1 and L3 SMGs of Atg5 WT mice, whereas the strongest ApopTag signals were detected in L3 and L7 SMGs of Atg5 KO mice. Magnification: × 100, and enlarged view ( inset ): × 400. Bar: 100 μ m. Percent ApopTag-positive cells quantified by dividing the total ApopTag-positive cells by total number of cells examined from 10 randomly chosen fields are shown ( right panel ). Student’s t -test was employed to determine statistically significant differences in percentage of ApopTag-positive cells between Atg5 WT and Atg5 KO groups. Results are shown as mean±S.D.; * P <0.05. ( b ) ATG5 deficiency delays caspase-3 activation. Equal amounts of whole SMG lysates from Ctrl, L1, L3 and L7 SMGs of Atg5 WT and Atg5 KO mice were analyzed on western blots using indicated antibodies. The cleavage of caspase-3 represents caspase-3 activation. ( c ) Gene expression analyses of Prol1/Muc-10 and Klk1 , markers for acinar and duct cells, respectively. SMG duct ligation was performed as described in . Expression of the indicated messages in ligated and control SMGs was analyzed by quantitative RT-PCR analyses. Relative gene expression was calculated where respective mean value for control SMG set to 1 for each amplification ( N ≥4). Non-parametric Mann–Whitney test was performed to compare expression levels between respective ligated and control SMGs ( *; below bars), and between same-day ligated SMGs and control SMGs from Atg5 WT and Atg5 KO ( *; above bars). Results are shown as mean±S.D.; * P <0.05; ** P <0.01. ( d ) Autophagy inhibition correlates with resistance to H 2 O 2 -induced cell death. Cell viability was measured in m5-7, Atg7 -KO MEF, WT MEF and salivary Pa-4 cells treated with indicated concentrations of H 2 O 2 in combination with chloroquine (CQ; 20 μ M) or baflomycin A1 (BafA1; 10 nM) for 48 h and compared with that of the control cells. Data were analyzed with ANOVA followed by Bonferroni t -test to determine the statistical differences between treatment and control group ( left panels ). Representative western blots ( right panels ), in which total lysates from cells treated for 6 h with vehicle, CQ (20 μ M) or BafA1 (10 nM) were stained with indicated antibodies, are shown. Results are shown as mean±S.E.M.; N =3; * P <0.05; ** P <0.01; *** P <0.001
Index in PubMed under a CC BY license. PMID: 25341032