PKCι/λ Antibody [H1H7] | Primary Antibodies

Application: Reactivity: Rat

Lane 1: Mouse brain

Experiment Essentials

WB
Recommended WB dilution ratio: 1:250

Usage Information

Dilution
WB1:250

Application
WB, IP, IHC, IF

Reactivity
Rat

Source
Mouse Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
68 kDa 74 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Positive Control	Rat cerebrum; Rat neurons
Negative Control

Experimental Methods

WB
Experimental Protocol: Sample preparation 1. Tissue: Lyse the tissue sample by adding an appropriate volume of ice-cold RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail)，and homogenize the tissue at a low temperature or lyse it by sonication on ice, then incubate on ice for 30 minutes. 2. Adherent cell: Aspirate the culture medium and wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) , sonicate to lyse the cells, and incubate on ice for 30 minutes. 3. Suspension cell: Transfer the culture medium to a pre-cooled centrifuge tube. Centrifuge and aspirate the supernatant. Wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) , sonicate to lyse the cells, and incubate on ice for 30 minutes. 4. Place the lysate into a pre-cooled microcentrifuge tube. Centrifuge at 4°C for 15 min. Collect the supernatant; 5. Remove a small volume of lysate to determine the protein concentration; 6. Combine the lysate with protein loading buffer. Boil 20 µL sample under 95-100°C for 5 min. Centrifuge for 5 min after cool down on ice. Electrophoretic separation 1. According to the concentration of extracted protein, load appropriate amount of protein sample and marker onto SDS-PAGE gels for electrophoresis. Recommended separating gel (lower gel) concentration: 10%. Reference Table for Selecting SDS-PAGE Separation Gel Concentrations 2. Power up 80V for 30 minutes. Then the power supply is adjusted (110 V~150 V), the Marker is observed, and the electrophoresis can be stopped when the indicator band of the predyed protein Marker where the protein is located is properly separated. (Note that the current should not be too large when electrophoresis, too large current (more than 150 mA) will cause the temperature to rise, affecting the result of running glue. If high currents cannot be avoided, an ice bath can be used to cool the bath.) Transfer membrane 1. Take out the converter, soak the clip and consumables in the pre-cooled converter; 2. Activate PVDF membrane with methanol for 1 min and rinse with transfer buffer; 3. Install it in the order of "black edge of clip - sponge - filter paper - filter paper - glue -PVDF membrane - filter paper - filter paper - sponge - white edge of clip"; 4. The protein was electrotransferred to PVDF membrane. ( 0.45 µm PVDF membrane is recommended ) Reference Table for Selecting PVDF Membrane Pore Size Specifications Recommended conditions for wet transfer: 200 mA, 120 min. ( Note that the transfer conditions can be adjusted according to the protein size. For high-molecular-weight proteins, a higher current and longer transfer time are recommended. However, ensure that the transfer tank remains at a low temperature to prevent gel melting.) Block 1. After electrotransfer, wash the film with TBST at room temperature for 5 minutes; 2. Incubate the film in the blocking solution for 1 hour at room temperature; 3. Wash the film with TBST for 3 times, 5 minutes each time. Antibody incubation 1. Use primary antibody dilution buffer to prepare the primary antibody working liquid (recommended dilution ratio for primary antibody 1:250), gently shake and incubate with the film at 4°C overnight; 2. Wash the film with TBST 3 times, 5 minutes each time; 3. Add the secondary antibody to the blocking solution and incubate with the film gently at room temperature for 1 hour; 4. After incubation, wash the film with TBST 3 times for 5 minutes each time. Antibody staining 1. Add the prepared ECL luminescent substrate (or select other color developing substrate according to the second antibody) and mix evenly; 2. Incubate with the film for 1 minute, remove excess substrate (keep the film moist), wrap with plastic film, and expose in the imaging system.

WB

Experimental Protocol:

Sample preparation

1. Tissue: Lyse the tissue sample by adding an appropriate volume of ice-cold RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail)，and homogenize the tissue at a low temperature or lyse it by sonication on ice, then incubate on ice for 30 minutes.

2. Adherent cell: Aspirate the culture medium and wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) , sonicate to lyse the cells, and incubate on ice for 30 minutes.

3. Suspension cell: Transfer the culture medium to a pre-cooled centrifuge tube. Centrifuge and aspirate the supernatant. Wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) , sonicate to lyse the cells, and incubate on ice for 30 minutes.

4. Place the lysate into a pre-cooled microcentrifuge tube. Centrifuge at 4°C for 15 min. Collect the supernatant;

5. Remove a small volume of lysate to determine the protein concentration;

6. Combine the lysate with protein loading buffer. Boil 20 µL sample under 95-100°C for 5 min. Centrifuge for 5 min after cool down on ice.

Electrophoretic separation

1. According to the concentration of extracted protein, load appropriate amount of protein sample and marker onto SDS-PAGE gels for electrophoresis. Recommended separating gel (lower gel) concentration: 10%. Reference Table for Selecting SDS-PAGE Separation Gel Concentrations

2. Power up 80V for 30 minutes. Then the power supply is adjusted (110 V~150 V), the Marker is observed, and the electrophoresis can be stopped when the indicator band of the predyed protein Marker where the protein is located is properly separated. (Note that the current should not be too large when electrophoresis, too large current (more than 150 mA) will cause the temperature to rise, affecting the result of running glue. If high currents cannot be avoided, an ice bath can be used to cool the bath.)

Transfer membrane

1. Take out the converter, soak the clip and consumables in the pre-cooled converter;

2. Activate PVDF membrane with methanol for 1 min and rinse with transfer buffer;

3. Install it in the order of "black edge of clip - sponge - filter paper - filter paper - glue -PVDF membrane - filter paper - filter paper - sponge - white edge of clip";

4. The protein was electrotransferred to PVDF membrane. ( 0.45 µm PVDF membrane is recommended ) Reference Table for Selecting PVDF Membrane Pore Size Specifications

Recommended conditions for wet transfer: 200 mA, 120 min.

( Note that the transfer conditions can be adjusted according to the protein size. For high-molecular-weight proteins, a higher current and longer transfer time are recommended. However, ensure that the transfer tank remains at a low temperature to prevent gel melting.)

Block

1. After electrotransfer, wash the film with TBST at room temperature for 5 minutes;

2. Incubate the film in the blocking solution for 1 hour at room temperature;

3. Wash the film with TBST for 3 times, 5 minutes each time.

Antibody incubation

1. Use primary antibody dilution buffer to prepare the primary antibody working liquid (recommended dilution ratio for primary antibody 1:250), gently shake and incubate with the film at 4°C overnight;

2. Wash the film with TBST 3 times, 5 minutes each time;

3. Add the secondary antibody to the blocking solution and incubate with the film gently at room temperature for 1 hour;

4. After incubation, wash the film with TBST 3 times for 5 minutes each time.

Antibody staining

1. Add the prepared ECL luminescent substrate (or select other color developing substrate according to the second antibody) and mix evenly;

2. Incubate with the film for 1 minute, remove excess substrate (keep the film moist), wrap with plastic film, and expose in the imaging system.

IF
Experimental Protocol: Sample Preparation 1. Adherent Cells: Place a clean, sterile coverslip in a culture dish. Once the cells grow to near confluence as a monolayer, remove the coverslip for further use. 2. Suspension Cells: Seed the cells onto a clean, sterile slide coated with poly-L-lysine. 3. Frozen Sections: Allow the slide to thaw at room temperature. Wash it with pure water or PBS for 2 times, 3 minutes each time. 4. Paraffin Sections: Deparaffinization and rehydration. Wash the slide with pure water or PBS for 3 times, 3 minutes each time. Then perform antigen retrieval. Fixation 1. Fix the cell coverslips/spots or tissue sections at room temperature using a fixative such as 4% paraformaldehyde (4% PFA) for 10-15 minutes. 2. Wash the sample with PBS for 3 times, 3 minutes each time. Permeabilization 1.Add a detergent such as 0.1–0.3% Triton X-100 to the sample and incubate at room temperature for 10–20 minutes. (Note: This step is only required for intracellular antigens. For antigens expressed on the cell membrane, this step is unnecessary.) Wash the sample with PBS for 3 times, 3 minutes each time. Blocking Add blocking solution and incubate at room temperature for at least 1 hour. (Common blocking solutions include: serum from the same source as the secondary antibody, BSA, or goat serum.) Note: Ensure the sample remains moist during and after the blocking step to prevent drying, which can lead to high background. Immunofluorescence Staining (Day 1) 1. Remove the blocking solution and add the diluted primary antibody. 2. Incubate the sample in a humidified chamber at 4°C overnight. Immunofluorescence Staining (Day 2) 1. Remove the primary antibody and wash with PBST for 3 times, 5 minutes each time. 2. Add the diluted fluorescent secondary antibody and incubate in the dark at 4°C for 1–2 hours. 3. Remove the secondary antibody and wash with PBST for 3 times, 5 minutes each time. 4. Add diluted DAPI and incubate at room temperature in the dark for 5–10 minutes. 5. Wash with PBST for 3 times, 5 minutes each time. Mounting 1. Mount the sample with an anti-fade mounting medium. 2. Allow the slide to dry at room temperature overnight in the dark. 3. Store the slide in a slide storage box at 4°C, protected from light.

IF

Experimental Protocol:

Sample Preparation

1. Adherent Cells: Place a clean, sterile coverslip in a culture dish. Once the cells grow to near confluence as a monolayer, remove the coverslip for further use.

2. Suspension Cells: Seed the cells onto a clean, sterile slide coated with poly-L-lysine.

3. Frozen Sections: Allow the slide to thaw at room temperature. Wash it with pure water or PBS for 2 times, 3 minutes each time.

4. Paraffin Sections: Deparaffinization and rehydration. Wash the slide with pure water or PBS for 3 times, 3 minutes each time. Then perform antigen retrieval.

Fixation

1. Fix the cell coverslips/spots or tissue sections at room temperature using a fixative such as 4% paraformaldehyde (4% PFA) for 10-15 minutes.

2. Wash the sample with PBS for 3 times, 3 minutes each time.

Permeabilization

1.Add a detergent such as 0.1–0.3% Triton X-100 to the sample and incubate at room temperature for 10–20 minutes.

(Note: This step is only required for intracellular antigens. For antigens expressed on the cell membrane, this step is unnecessary.)

Wash the sample with PBS for 3 times, 3 minutes each time.

Blocking

Add blocking solution and incubate at room temperature for at least 1 hour. (Common blocking solutions include: serum from the same source as the secondary antibody, BSA, or goat serum.)

Note: Ensure the sample remains moist during and after the blocking step to prevent drying, which can lead to high background.

Immunofluorescence Staining (Day 1)

1. Remove the blocking solution and add the diluted primary antibody.

2. Incubate the sample in a humidified chamber at 4°C overnight.

Immunofluorescence Staining (Day 2)

1. Remove the primary antibody and wash with PBST for 3 times, 5 minutes each time.

2. Add the diluted fluorescent secondary antibody and incubate in the dark at 4°C for 1–2 hours.

3. Remove the secondary antibody and wash with PBST for 3 times, 5 minutes each time.

4. Add diluted DAPI and incubate at room temperature in the dark for 5–10 minutes.

5. Wash with PBST for 3 times, 5 minutes each time.

Mounting

1. Mount the sample with an anti-fade mounting medium.

2. Allow the slide to dry at room temperature overnight in the dark.

3. Store the slide in a slide storage box at 4°C, protected from light.

IHC
Experimental Protocol： Deparaffinization/Rehydration 1. Deparaffinize/hydrate sections: 2. Incubate sections in three washes of xylene for 5 min each. 3. Incubate sections in two washes of 100% ethanol for 10 min each. 4. Incubate sections in two washes of 95% ethanol for 10 min each. 5. Wash sections two times in dH2O for 5 min each. 6.Antigen retrieval: For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; continue with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min. Staining 1. Wash sections in dH2O three times for 5 min each. 2. Incubate sections in 3% hydrogen peroxide for 10 min. 3. Wash sections in dH2O two times for 5 min each. 4. Wash sections in wash buffer for 5 min. 5. Block each section with 100–400 µl of blocking solution for 1 hr at room temperature. 6. Remove blocking solution and add 100–400 µl primary antibody diluent in to each section. Incubate overnight at 4°C. 7. Remove antibody solution and wash sections with wash buffer three times for 5 min each. 8. Cover section with 1–3 drops HRPas needed. Incubate in a humidified chamber for 30 min at room temperature. 9. Wash sections three times with wash buffer for 5 min each. 10. Add DAB Chromogen Concentrate to DAB Diluent and mix well before use. 11. Apply 100–400 µl DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity. 12. Immerse slides in dH2O. 13. If desired, counterstain sections with hematoxylin. 14. Wash sections in dH2O two times for 5 min each. 15. Dehydrate sections: Incubate sections in 95% ethanol two times for 10 sec each; Repeat in 100% ethanol, incubating sections two times for 10 sec each; Repeat in xylene, incubating sections two times for 10 sec each. 16. Mount sections with coverslips and mounting medium.

IHC

Experimental Protocol：

Deparaffinization/Rehydration

1. Deparaffinize/hydrate sections:

2. Incubate sections in three washes of xylene for 5 min each.

3. Incubate sections in two washes of 100% ethanol for 10 min each.

4. Incubate sections in two washes of 95% ethanol for 10 min each.

5. Wash sections two times in dH2O for 5 min each.

6.Antigen retrieval: For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; continue with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.

Staining

1. Wash sections in dH2O three times for 5 min each.

2. Incubate sections in 3% hydrogen peroxide for 10 min.

3. Wash sections in dH2O two times for 5 min each.

4. Wash sections in wash buffer for 5 min.

5. Block each section with 100–400 µl of blocking solution for 1 hr at room temperature.

6. Remove blocking solution and add 100–400 µl primary antibody diluent in to each section. Incubate overnight at 4°C.

7. Remove antibody solution and wash sections with wash buffer three times for 5 min each.

8. Cover section with 1–3 drops HRPas needed. Incubate in a humidified chamber for 30 min at room temperature.

9. Wash sections three times with wash buffer for 5 min each.

10. Add DAB Chromogen Concentrate to DAB Diluent and mix well before use.

11. Apply 100–400 µl DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity.

12. Immerse slides in dH2O.

13. If desired, counterstain sections with hematoxylin.

14. Wash sections in dH2O two times for 5 min each.

15. Dehydrate sections: Incubate sections in 95% ethanol two times for 10 sec each; Repeat in 100% ethanol, incubating sections two times for 10 sec each; Repeat in xylene, incubating sections two times for 10 sec each.

16. Mount sections with coverslips and mounting medium.

Datasheet & SDS

Biological Description

Specificity
PKCι/λ Antibody (Mouse mAb) [H1H7] detects endogenous levels of total PKCι/λ protein.

Subcellular Location
Cytoplasm, Endosome, Membrane, Nucleus

Uniprot ID
P41743

Clone
H1H7

Synonym(s)
Protein kinase C iota type, Atypical protein kinase C-lambda/iota (PRKC-lambda/iota; aPKC-lambda/iota) nPKC-iota, PRKCI, DXS1179E

Background
Protein kinase C iota (PKCι), also designated PKCλ in mice, belongs to the atypical protein kinase C (aPKC) subfamily comprising PKCι and PKCζ, which exhibit calcium and diacylglycerol independence distinguishing them from conventional and novel PKC isoforms. PKCι contains an N-terminal regulatory domain featuring a single atypical cysteine-rich C1 domain lacking diacylglycerol binding capability, a Phox/Bem1 (PB1) domain mediating protein-protein interactions, and a pseudosubstrate motif, alongside a C-terminal catalytic serine/threonine kinase domain activated through phosphorylation at Thr500 in the activation loop, Thr641 via autophosphorylation, and Ser660 at the hydrophobic motif by PDK1 or related kinases. The protein exhibits ubiquitous tissue expression with particularly high levels in brain, lung, liver, and kidney, contrasting with the restricted expression pattern of PKCζ, and demonstrates 72% overall amino acid sequence identity to PKCζ despite exhibiting distinct, non-redundant cellular functions as evidenced by differential embryonic lethality upon genetic disruption. PKCι functions as a critical oncogene required for transformed growth, invasion, chemoresistance, and tumor cell survival across multiple human malignancies including non-small cell lung cancer, ovarian carcinoma, pancreatic adenocarcinoma, prostate cancer, colon carcinoma, and glioblastoma, with the PKCι gene on chromosome 3q26 representing a frequent target of tumor-specific gene amplification in approximately 36% of NSCLC cases and 70% of lung squamous cell carcinomas. PKCι drives oncogenic signaling through formation of a core complex with the scaffold protein Par6 and the small GTPase Rac1 or Cdc42, wherein the PB1 domain of PKCι heterodimerizes with the PB1 domain of Par6 while Par6's CRIB motif binds GTP-loaded Rac1 or Cdc42, coupling PKCι to polarity establishment, cytoskeletal reorganization, and transformed growth pathways. The PKCι-Par6-Rac1 polarity complex activates a Rac1→p21-activated kinase (PAK)→MEK→ERK signaling cascade essential for anchorage-independent transformed growth and tumorigenicity, with expression of kinase-deficient PKCι or constitutive disruption of the PKCι-Par6 interaction abolishing soft agar colony formation and tumor development without affecting adherent cell growth. PKCι promotes cell survival through at least three distinct mechanisms depending on cellular context—in chronic myelogenous leukemia cells, PKCι activates both canonical and non-canonical NF-κB signaling to induce IκB degradation and directly trans-activate nuclear NF-κB transcriptional activity conferring chemoresistance, in NSCLC cells Src-activated PKCι directly phosphorylates the pro-apoptotic protein Bad preventing its interaction with Bcl-XL to enhance survival, and in glioblastoma PKCι attenuates p38 MAPK signaling to reduce cisplatin-induced apoptosis. The protein functions downstream of oncogenic Ras, with Ras directly binding and activating PKCι, and PKCι proving essential for Ras-mediated transformation, invasion, and anchorage-independent growth in intestinal epithelial cells and critical for K-Ras-dependent colon carcinogenesis and lung tumor development. PKCι mediates cellular invasion through direct phosphorylation of μ- and m-calpains promoting wound healing and migration in NSCLC cells, and operates within a PKCβII→Ras→PKCι/Rac1→MEK signaling pathway driving invasive phenotypes in colon cancer cells. The PB1 domain contains a unique cysteine residue at position 69 (Cys69) located within the conserved OPCA motif at the PKCι-Par6 binding interface, where it interacts with Arg28 of Par6, and this cysteine serves as the molecular target for the selective PKCι inhibitor aurothiomalate (ATM) which forms a gold-cysteine adduct disrupting PKCι-Par6 and PKCι-p62 interactions without affecting other PB1 domain interactions. PKCι expression correlates with poor clinical outcome in NSCLC patients, with elevated PKCι conferring a 2.6-fold increased risk of cancer-related death and early-stage NSCLC patients with high PKCι exhibiting an 11-fold increased risk of relapse compared to those with low PKCι expression, positioning PKCι as both a prognostic biomarker and therapeutic target. Bcr-Abl transcriptionally activates PKCι through Ras/MEK-dependent induction of an Elk1 element within the proximal PKCι promoter, representing one mechanism of PKCι upregulation beyond gene amplification, although somatic mutations in PKCι appear absent or extremely rare based on sequencing analysis of all 18 exons in NSCLC cases.

Background

Protein kinase C iota (PKCι), also designated PKCλ in mice, belongs to the atypical protein kinase C (aPKC) subfamily comprising PKCι and PKCζ, which exhibit calcium and diacylglycerol independence distinguishing them from conventional and novel PKC isoforms. PKCι contains an N-terminal regulatory domain featuring a single atypical cysteine-rich C1 domain lacking diacylglycerol binding capability, a Phox/Bem1 (PB1) domain mediating protein-protein interactions, and a pseudosubstrate motif, alongside a C-terminal catalytic serine/threonine kinase domain activated through phosphorylation at Thr500 in the activation loop, Thr641 via autophosphorylation, and Ser660 at the hydrophobic motif by PDK1 or related kinases. The protein exhibits ubiquitous tissue expression with particularly high levels in brain, lung, liver, and kidney, contrasting with the restricted expression pattern of PKCζ, and demonstrates 72% overall amino acid sequence identity to PKCζ despite exhibiting distinct, non-redundant cellular functions as evidenced by differential embryonic lethality upon genetic disruption. PKCι functions as a critical oncogene required for transformed growth, invasion, chemoresistance, and tumor cell survival across multiple human malignancies including non-small cell lung cancer, ovarian carcinoma, pancreatic adenocarcinoma, prostate cancer, colon carcinoma, and glioblastoma, with the PKCι gene on chromosome 3q26 representing a frequent target of tumor-specific gene amplification in approximately 36% of NSCLC cases and 70% of lung squamous cell carcinomas. PKCι drives oncogenic signaling through formation of a core complex with the scaffold protein Par6 and the small GTPase Rac1 or Cdc42, wherein the PB1 domain of PKCι heterodimerizes with the PB1 domain of Par6 while Par6's CRIB motif binds GTP-loaded Rac1 or Cdc42, coupling PKCι to polarity establishment, cytoskeletal reorganization, and transformed growth pathways. The PKCι-Par6-Rac1 polarity complex activates a Rac1→p21-activated kinase (PAK)→MEK→ERK signaling cascade essential for anchorage-independent transformed growth and tumorigenicity, with expression of kinase-deficient PKCι or constitutive disruption of the PKCι-Par6 interaction abolishing soft agar colony formation and tumor development without affecting adherent cell growth. PKCι promotes cell survival through at least three distinct mechanisms depending on cellular context—in chronic myelogenous leukemia cells, PKCι activates both canonical and non-canonical NF-κB signaling to induce IκB degradation and directly trans-activate nuclear NF-κB transcriptional activity conferring chemoresistance, in NSCLC cells Src-activated PKCι directly phosphorylates the pro-apoptotic protein Bad preventing its interaction with Bcl-XL to enhance survival, and in glioblastoma PKCι attenuates p38 MAPK signaling to reduce cisplatin-induced apoptosis. The protein functions downstream of oncogenic Ras, with Ras directly binding and activating PKCι, and PKCι proving essential for Ras-mediated transformation, invasion, and anchorage-independent growth in intestinal epithelial cells and critical for K-Ras-dependent colon carcinogenesis and lung tumor development. PKCι mediates cellular invasion through direct phosphorylation of μ- and m-calpains promoting wound healing and migration in NSCLC cells, and operates within a PKCβII→Ras→PKCι/Rac1→MEK signaling pathway driving invasive phenotypes in colon cancer cells. The PB1 domain contains a unique cysteine residue at position 69 (Cys69) located within the conserved OPCA motif at the PKCι-Par6 binding interface, where it interacts with Arg28 of Par6, and this cysteine serves as the molecular target for the selective PKCι inhibitor aurothiomalate (ATM) which forms a gold-cysteine adduct disrupting PKCι-Par6 and PKCι-p62 interactions without affecting other PB1 domain interactions. PKCι expression correlates with poor clinical outcome in NSCLC patients, with elevated PKCι conferring a 2.6-fold increased risk of cancer-related death and early-stage NSCLC patients with high PKCι exhibiting an 11-fold increased risk of relapse compared to those with low PKCι expression, positioning PKCι as both a prognostic biomarker and therapeutic target. Bcr-Abl transcriptionally activates PKCι through Ras/MEK-dependent induction of an Elk1 element within the proximal PKCι promoter, representing one mechanism of PKCι upregulation beyond gene amplification, although somatic mutations in PKCι appear absent or extremely rare based on sequencing analysis of all 18 exons in NSCLC cases.

References
https://pubmed.ncbi.nlm.nih.gov/17570678/ https://pubmed.ncbi.nlm.nih.gov/20945390/

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%

PKCι/λ Antibody (Mouse mAb) [H1H7]