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Pan-Lactyl-lysine Recombinant Rabbit Monoclonal Antibody [PSH03-74] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750908

Pan-Lactyl-lysine Recombinant Rabbit Monoclonal Antibody [PSH03-74] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IF-Cell

  • Dot Blot

Reactivity

  • Species independent

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

Pan-Lactyl-lysine Recombinant Rabbit Monoclonal Antibody [PSH03-74] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic Lactyl lysine-containing peptide.

Species Reactivity

Species independent

Validated Applications

WB, IHC-P, IF-Cell, Dot Blot

Positive Control

HeLa treated with 100mM Lactate sodium for 24 hours cell lysate, human breast cancer tissue, human colon cancer tissue, mouse small intestine tissue, rat small intestine tissue, HeLa cells treated with 100mM Lactate sodium for 24 hours.

Conjugation

unconjugated

Clone Number

PSH03-74

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:500-1:1,000

  • IF-Cell

  • 1:1,000

  • Dot Blot

  • 1:2,000

Target

Function

Post-translational modifications (PTMs) represent a crucial means of regulating diverse biological processes and cellular physiology by influencing protein structure and function. Lactate, as an important metabolite in the body, plays an important role in both physiological and pathological processes. The level of lactylation is closely related to the abundance of lactate. The Warburg effect was originally used to describe a phenomenon of increased lactate production in tumors, which is involved in the regulation of various cellular processes such as angiogenesis, hypoxia, macrophage polarization, and T cell activation. Lactylation has been reported to be closely related to the occurrence and development of various diseases, such as tumor formation, sepsis, autoimmune diseases, and neurodegenerative diseases. The main modification enzymes (writers) and erasers (erasers) of lactylation are P300 and HDAC1-3, respectively. As a large number of studies have demonstrated that lysine acylation possessed a diverse range of substrate proteins, however, no systematic analysis has been reported for lysine lactylation.

Background References

1. Walsh, C. T., Garneau-Tsodikova, S., and Gatto, G. J. Jr. (2005). Protein posttranslational modifications: the chemistry of proteome diversifications. Angew. Chem. Int. Ed. Engl. 44, 7342–7372.

2. Zhang, D., Tang, Z., Huang, H., Zhou, G., Cui, C., Hu, R., et al. (2019). Metabolic regulation of gene expression by histone lactylation. Nature 574, 575–580.

Synonyms

Lactyllysine antibody

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Pan-Lactyl-lysine on different lysates with Rabbit anti-Pan-Lactyl-lysine antibody (<a href="/products/HA750908" style="font-weight: bold;text-decoration: underline;">HA750908</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 100mM Lactate sodium for 24 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Observed band size: Mutiple kDa<br /><br />Exposure time: 59 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750908" style="font-weight: bold;text-decoration: underline;">HA750908</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Pan-Lactyl-lysine on different lysates with Rabbit anti-Pan-Lactyl-lysine antibody (HA750908) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 100mM Lactate sodium for 24 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

    Observed band size: Mutiple kDa

    Exposure time: 59 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750908) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of HeLa cells treated with or without 100mM Lactate sodium for 24 hours labeling Pan-Lactyl-lysine with Rabbit anti-Pan-Lactyl-lysine antibody (<a href="/products/HA750908" style="font-weight: bold;text-decoration: underline;">HA750908</a>) at 1/1,000 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Pan-Lactyl-lysine antibody (<a href="/products/HA750908" style="font-weight: bold;text-decoration: underline;">HA750908</a>) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of HeLa cells treated with or without 100mM Lactate sodium for 24 hours labeling Pan-Lactyl-lysine with Rabbit anti-Pan-Lactyl-lysine antibody (HA750908) at 1/1,000 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Pan-Lactyl-lysine antibody (HA750908) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Pan-Lactyl-lysine antibody (<a href="/products/HA750908" style="font-weight: bold;text-decoration: underline;">HA750908</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750908" style="font-weight: bold;text-decoration: underline;">HA750908</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Pan-Lactyl-lysine antibody (HA750908) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750908) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Pan-Lactyl-lysine antibody (<a href="/products/HA750908" style="font-weight: bold;text-decoration: underline;">HA750908</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750908" style="font-weight: bold;text-decoration: underline;">HA750908</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Pan-Lactyl-lysine antibody (HA750908) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750908) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-Pan-Lactyl-lysine antibody (<a href="/products/HA750908" style="font-weight: bold;text-decoration: underline;">HA750908</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750908" style="font-weight: bold;text-decoration: underline;">HA750908</a>) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-Pan-Lactyl-lysine antibody (HA750908) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750908) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Rabbit anti-Pan-Lactyl-lysine antibody (<a href="/products/HA750908" style="font-weight: bold;text-decoration: underline;">HA750908</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750908" style="font-weight: bold;text-decoration: underline;">HA750908</a>) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Rabbit anti-Pan-Lactyl-lysine antibody (HA750908) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750908) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Dot blot analysis of Pan-Lactyl-lysine on different proteins with Rabbit anti-Pan-Lactyl-lysine antibody (<a href="/products/HA750908" style="font-weight: bold;text-decoration: underline;">HA750908</a>) at 1/2,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution for 1 hour at room temperature.<br /><br />Lane 1: Lac pepetide library (positive)<br />Lane 2: Ac peptide library (negative)<br />Lane 3: Non-lac peptide library (negative)<br /><br />Proteins loading: 100ng, 20ng, 4ng;<br /><br />Blocking and dilution buffer: 5% NFDM/TBST;<br /><br />Exposure time: 20 seconds.

    Dot blot analysis of Pan-Lactyl-lysine on different proteins with Rabbit anti-Pan-Lactyl-lysine antibody (HA750908) at 1/2,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution for 1 hour at room temperature.

    Lane 1: Lac pepetide library (positive)
    Lane 2: Ac peptide library (negative)
    Lane 3: Non-lac peptide library (negative)

    Proteins loading: 100ng, 20ng, 4ng;

    Blocking and dilution buffer: 5% NFDM/TBST;

    Exposure time: 20 seconds.

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