PKM1 Antibody - BSA Free

Novus Biologicals | Catalog # NBP2-14833

Novus Biologicals
100% Guarantee

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Bovine

Cited:

Human, Mouse, Bovine

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunoblotting, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all PKM1 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to a C-terminal portion of the human PKM1 protein (between residues 350-450) [UniProt P14618-2].

Reactivity Notes

Bovine reactivity reported in scientific literature (PMID: 25416385).

Localization

Cytoplasm.

Specificity

This antibody is specific for the M1 isoform of PKM.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for PKM1 Antibody - BSA Free

Western Blot: PKM1 AntibodyBSA Free [NBP2-14833]

Western Blot: PKM1 AntibodyBSA Free [NBP2-14833]

PKM1-Antibody-Western-Blot-NBP2-14833-img0008.jpg
Immunohistochemistry-Paraffin: PKM1 Antibody - BSA Free [NBP2-14833]

Immunohistochemistry-Paraffin: PKM1 Antibody - BSA Free [NBP2-14833]

Immunohistochemistry-Paraffin: PKM1 Antibody [NBP2-14833] - PKM1 antibody was tested in human breast cancer using DAB with hematoxylin counterstain.
Western Blot: PKM1 AntibodyBSA Free [NBP2-14833]

Western Blot: PKM1 AntibodyBSA Free [NBP2-14833]

Western Blot: PKM1 Antibody [NBP2-14833] - PKM1 antibody was tested in 1. human skeletal muscle 2. mouse skeletal muscle 3. human brain 4. mouse brain and 5. human heart cell lysate.
Western Blot: PKM1 AntibodyBSA Free [NBP2-14833]

Western Blot: PKM1 AntibodyBSA Free [NBP2-14833]

Western Blot: PKM1 Antibody [NBP2-14833] - Detection of PKM1 on HEK293T and U-87 cells. Lane 1: HEK293T cells (major isoform is PKM2, human). Lane 2: HEK293T cells with plasmid overexpressing mouse PKM1. Lane 3: HEK293T cells with plasmid overexpressing mouse PKM2. Lane 4: U-87 cells with control shRNA (major isoform is PKM2, human). Lane 5: U-87 cells with PTB/A1/A2 shRNAs (major isoform is PKM1, human). Photo courtesy of product review by verified customer.
Western Blot: PKM1 AntibodyBSA Free [NBP2-14833]

Western Blot: PKM1 AntibodyBSA Free [NBP2-14833]

PKM1-Antibody-Western-Blot-NBP2-14833-img0006.jpg
Western Blot: PKM1 AntibodyBSA Free [NBP2-14833]

Western Blot: PKM1 AntibodyBSA Free [NBP2-14833]

PKM1-Antibody-Western-Blot-NBP2-14833-img0007.jpg
Simple Western: PKM1 AntibodyBSA Free [NBP2-14833]

Simple Western: PKM1 AntibodyBSA Free [NBP2-14833]

Simple Western: PKM1 Antibody [NBP2-14833] - Simple Western lane view shows a specific band for PKM1 in 0.5 mg/ml of Human Brain lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
PKM1 Antibody - BSA Free

Immunohistochemistry-Paraffin: PKM1 Antibody - BSA Free [NBP2-14833] -

Immunohistochemistry-Paraffin: PKM1 Antibody - BSA Free [NBP2-14833] - Expression of PKM1 & PKM2 in clinical colorectal cancer samples.(a) The protein expression of PKM1 & PKM2 in clinical specimens of cancer tumor (T) & the adjacent normal tissues (N) is shown. PKM1 & PKM2 were detected by Western blotting in under the same experimental conditions at the same time. The full-length blots are presented in Supplementary Figure S3b. (b–d) Immunohistochemical staining of normal colon tissue adjacent to tumor tissue of case 10. Results of H&E staining (b), staining with anti-PKM1 antibody (c), & staining with anti-PKM2 (d) are shown. The boxed regions in “c” & “d” are enlarged in the images below. (e–h) Immunohistochemical staining of clinical colorectal cancer tissue specimen of representative case 3. H&E-stained section with normal tissue (upper right corner) neighboring the tumor area in the section is shown (e), along with the same section stained with anti-PKM2 antibody (f). Enlarged views of boxed areas in “f” show normal colorectal crypt in mucosa (g) & tumor area (h) stained with anti-PKM2 antibody. Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/srep08647), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
PKM1 Antibody - BSA Free

Western Blot: PKM1 Antibody - BSA Free [NBP2-14833] -

Western Blot: PKM1 Antibody - BSA Free [NBP2-14833] - (a) Luciferase activities after co-transfection of DLD-1 cells w/ control or miR-124 (wild-type or mutant-type) pMIR vectors having predictive miR-124 binding site in 3′UTR of PTB1. Upper panel region of 3′-UTR of human PTB1 mRNA complementary to mature miR-124. Box indicates predicted binding sites for miR-124. (b) Same as “a” except miR-133b used. (c) Expression of PTB1, PKM1, & PKM2 proteins at 72 h after transfection of DLD-1, NB9 or IMR-32 cells w/ miR-124 (10, 20 or 40 nM). (d) Expression of PTB1, PKM1, & PKM2 proteins at 72 h after transfection of DLD-1, RD or KYM-1 cells w/ miR-133b (10, 20 nM). (e) Expression of PTB1, PKM1, & PKM2 proteins at 72 h after transfection of DLD-1, NB-9 or RD cells w/ siR-PTB1 (2, 5 nM). (f) Effect of combined treatment of DLD-1 cells w/ antagomiR-124 & miR-124 or antagomiR-133b & miR-133b. DLD-1 cells transfected w/ non-specific control, miR-124/miR-133b (10 nM), miR-124/miR-133b (10 nM) + antagomiR-124/antagomiR-133b (5 nM) or miR-124/miR-133b (10 nM) + antagomiR-124/antagomiR-133b (10 nM). Expression level of PTB1 assessed at 48 h after transfection. The full-length blots are presented in Supplementary Figure S3a. (g) IF of PKM1 (upper panels) & PKM2 (lower panels) at 48 h after transfection of DLD-1 cells w/ miR-124 (20 nM) or miR-133b (20 nM). Left panels, treatment w/ control miRNA; middle panels, treatment w/ miR-124; right panels, treatment w/ miR-133b. PKM1 or PKM2 is stained red, & nuclei are stained blue. (h) Lactate production measured at 48 h after transfection of DLD-1 cells w/ miR-124 (20 nM), miR-133b (20 nM) or siR-PTB1 (5 nM). Results are presented as mean± SD (* P < 0.05; ** P < 0.01; *** P < 0.001; N.S., not statistically significant). Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/srep08647), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
PKM1 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: PKM1 Antibody - BSA Free [NBP2-14833] -

Immunocytochemistry/ Immunofluorescence: PKM1 Antibody - BSA Free [NBP2-14833] - (a) Luciferase activities after co-transfection of DLD-1 cells w/ control or miR-124 (wild-type or mutant-type) pMIR vectors having predictive miR-124 binding site in 3′UTR of PTB1. Upper panel region of 3′-UTR of human PTB1 mRNA complementary to mature miR-124. Box indicates predicted binding sites for miR-124. (b) Same as “a” except miR-133b used. (c) Expression of PTB1, PKM1, & PKM2 proteins at 72 h after transfection of DLD-1, NB9 or IMR-32 cells w/ miR-124 (10, 20 or 40 nM). (d) Expression of PTB1, PKM1, & PKM2 proteins at 72 h after transfection of DLD-1, RD or KYM-1 cells w/ miR-133b (10, 20 nM). (e) Expression of PTB1, PKM1, & PKM2 proteins at 72 h after transfection of DLD-1, NB-9 or RD cells w/ siR-PTB1 (2, 5 nM). (f) Effect of combined treatment of DLD-1 cells w/ antagomiR-124 & miR-124 or antagomiR-133b & miR-133b. DLD-1 cells transfected w/ non-specific control, miR-124/miR-133b (10 nM), miR-124/miR-133b (10 nM) + antagomiR-124/antagomiR-133b (5 nM) or miR-124/miR-133b (10 nM) + antagomiR-124/antagomiR-133b (10 nM). Expression level of PTB1 assessed at 48 h after transfection. The full-length blots are presented in Supplementary Figure S3a. (g) IF of PKM1 (upper panels) & PKM2 (lower panels) at 48 h after transfection of DLD-1 cells w/ miR-124 (20 nM) or miR-133b (20 nM). Left panels, treatment w/ control miRNA; middle panels, treatment w/ miR-124; right panels, treatment w/ miR-133b. PKM1 or PKM2 is stained red, & nuclei are stained blue. (h) Lactate production measured at 48 h after transfection of DLD-1 cells w/ miR-124 (20 nM), miR-133b (20 nM) or siR-PTB1 (5 nM). Results are presented as mean± SD (* P < 0.05; ** P < 0.01; *** P < 0.001; N.S., not statistically significant). Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/srep08647), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
PKM1 Antibody - BSA Free

Western Blot: PKM1 Antibody - BSA Free [NBP2-14833] -

Western Blot: PKM1 Antibody - BSA Free [NBP2-14833] - (a) Luciferase activities after co-transfection of DLD-1 cells w/ control or miR-124 (wild-type or mutant-type) pMIR vectors having predictive miR-124 binding site in 3′UTR of PTB1. Upper panel region of 3′-UTR of human PTB1 mRNA complementary to mature miR-124. Box indicates predicted binding sites for miR-124. (b) Same as “a” except miR-133b used. (c) Expression of PTB1, PKM1, & PKM2 proteins at 72 h after transfection of DLD-1, NB9 or IMR-32 cells w/ miR-124 (10, 20 or 40 nM). (d) Expression of PTB1, PKM1, & PKM2 proteins at 72 h after transfection of DLD-1, RD or KYM-1 cells w/ miR-133b (10, 20 nM). (e) Expression of PTB1, PKM1, & PKM2 proteins at 72 h after transfection of DLD-1, NB-9 or RD cells w/ siR-PTB1 (2, 5 nM). (f) Effect of combined treatment of DLD-1 cells w/ antagomiR-124 & miR-124 or antagomiR-133b & miR-133b. DLD-1 cells transfected w/ non-specific control, miR-124/miR-133b (10 nM), miR-124/miR-133b (10 nM) + antagomiR-124/antagomiR-133b (5 nM) or miR-124/miR-133b (10 nM) + antagomiR-124/antagomiR-133b (10 nM). Expression level of PTB1 assessed at 48 h after transfection. The full-length blots are presented in Supplementary Figure S3a. (g) IF of PKM1 (upper panels) & PKM2 (lower panels) at 48 h after transfection of DLD-1 cells w/ miR-124 (20 nM) or miR-133b (20 nM). Left panels, treatment w/ control miRNA; middle panels, treatment w/ miR-124; right panels, treatment w/ miR-133b. PKM1 or PKM2 is stained red, & nuclei are stained blue. (h) Lactate production measured at 48 h after transfection of DLD-1 cells w/ miR-124 (20 nM), miR-133b (20 nM) or siR-PTB1 (5 nM). Results are presented as mean± SD (* P < 0.05; ** P < 0.01; *** P < 0.001; N.S., not statistically significant). Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/srep08647), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
PKM1 Antibody - BSA Free

Western Blot: PKM1 Antibody - BSA Free [NBP2-14833] -

Western Blot: PKM1 Antibody - BSA Free [NBP2-14833] - Expression of PKM1 & PKM2 in clinical colorectal cancer samples.(a) The protein expression of PKM1 & PKM2 in clinical specimens of cancer tumor (T) & the adjacent normal tissues (N) is shown. PKM1 & PKM2 were detected by Western blotting in under the same experimental conditions at the same time. The full-length blots are presented in Supplementary Figure S3b. (b–d) Immunohistochemical staining of normal colon tissue adjacent to tumor tissue of case 10. Results of H&E staining (b), staining with anti-PKM1 antibody (c), & staining with anti-PKM2 (d) are shown. The boxed regions in “c” & “d” are enlarged in the images below. (e–h) Immunohistochemical staining of clinical colorectal cancer tissue specimen of representative case 3. H&E-stained section with normal tissue (upper right corner) neighboring the tumor area in the section is shown (e), along with the same section stained with anti-PKM2 antibody (f). Enlarged views of boxed areas in “f” show normal colorectal crypt in mucosa (g) & tumor area (h) stained with anti-PKM2 antibody. Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/srep08647), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
PKM1 Antibody - BSA Free

Western Blot: PKM1 Antibody - BSA Free [NBP2-14833] -

Western Blot: PKM1 Antibody - BSA Free [NBP2-14833] - Characterization of NOX4 & PKM2 in human RCC tumors & adjacent tissue. a Mitochondrial fractions were prepared from human tumors (T) or uninvolved adjacent tissue (N). NOX4 expression was examined by western blot analysis. Prohibitin was probed as a mitochondrial marker & loading control. b Quantitation of NOX4 distribution in the mitochondrial fraction from a. The results are expressed as the means using one-way ANOVA with Tukey’s post hoc test where ± S.E.M. *p < 0.05 compared to normal (N). c Mitochondria fractions were prepared from RCC tumors & NADPH-dependent superoxide generation was examined in the presence (+) or absence (−) of ATP. The results are from eight tumors & are expressed as the means using one-way ANOVA with Tukey’s post hoc test where ±S.E.M. **p < 0.01 is compared to without (−) ATP. d PKM2 & PKM1 expression was examined by western blot analysis in lysates prepared from human tumors (T) or uninvolved adjacent tissue (N) from the same patient. Actin as loading control Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29051480), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for PKM1 Antibody - BSA Free

Application
Recommended Usage

Immunoblotting

reported in scientific literature (PMID 25416385)

Immunocytochemistry/ Immunofluorescence

reported in multiple pieces of scientific literature

Immunohistochemistry

1:400

Immunohistochemistry-Paraffin

1:400

Simple Western

1:400

Western Blot

0.5 ug/ml
Application Notes
In Western blot a band is seen ~57 kDa in human and mouse skeletal muscle, brain and heart. In IHC-P, staining was observed in the cytoplasm of human breast cancer tissue. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in Human Brain lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:400, apparent MW was 59 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 3 reviews rated 5 using NBP2-14833 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 30% Glycerol

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.17 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: PKM1

Pyruvate kinases are glycolytic enzymes that transfer a phosphate group from phosphoenolpyruvate to ADP generating pyruvate and ATP. There are 4 isozymes of pyruvate kinase in mammals: L, R, M1 and M2. L type is major isozyme in the liver, R is found in red cells, M1 is the main form in muscle, heart and brain, and M2 is found in early fetal tissues as well as in most cancer cells. The M1 isoform differs from the M2 isoform due to a change in sequence between residues 389-433. In the presence of D-fructose 1,6-bisphosphate (FBP) the monomer reversibly forms the highly active tetrameric form. This tetramer is associated with the glycolytic enzyme complex and activates the pyruvate kinase activity. A nearly inactive dimeric form is associated with tumor cells.

Alternate Names

PK1, Pyruvate kinase isozymes M1, Pyruvate kinase M1, Pyruvate kinase isozyme M1

Entrez Gene IDs

5315 (Human); 18746 (Mouse)

Gene Symbol

PKM

UniProt

P14618 (Human), P52480 (Mouse)

Additional PKM1 Products

Product Documents for PKM1 Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for PKM1 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for PKM1 Antibody - BSA Free

Customer Reviews for PKM1 Antibody - BSA Free (3)

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  • PKM1 Antibody
    Name: Min-Joon Han
    Application: Western Blot
    Sample Tested: Human fibroblast
    Species: Human
    Verified Customer | Posted 10/24/2018
    PKM1 Antibody - BSA Free NBP2-14833
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: HCT116 cell line lysate
    Species: Human
    Verified Customer | Posted 03/31/2016
  • Name: Corey.
    Application: Western Blot
    Sample Tested: U-87 cells with PTB/A1/A2 shRNAs
    Species: Human
    Verified Customer | Posted 12/10/2012
    PKM1 Antibody - BSA Free NBP2-14833

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Protocols

View specific protocols for PKM1 Antibody - BSA Free (NBP2-14833):


Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 25 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

*Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for PKM1 Antibody - BSA Free

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  • Q: I just wanted to make sure the L/R didn't hit M1 and M2. I do not need to distinguish between L and R. So do you have an L/R specific monoclonal antibody for western blot?

    A: We do have an antibody that is specific for PKM1, however it is a polyclonal. The catalog number is NBP2-14833.

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