Lactate Dehydrogenase A/LDHA Antibody - BSA Free

Novus Biologicals | Catalog # NBP1-48336

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Porcine, Bovine

Cited:

Human, Mouse, Rat, Bovine

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

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

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all Lactate Dehydrogenase A/LDHA Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide made to a C-terminal portion of the human Lactate Dehydrogenase A protein (within residues 280-332). [Swiss-Prot# P00338]

Reactivity Notes

Porcine reactivity reported in scientific literature (Elgin et al).

Localization

Cytoplasm.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

36 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Lactate Dehydrogenase A/LDHA Antibody - BSA Free

Western Blot: Lactate Dehydrogenase A/LDHA AntibodyBSA Free [NBP1-48336]

Western Blot: Lactate Dehydrogenase A/LDHA AntibodyBSA Free [NBP1-48336]

Lactate-Dehydrogenase-A-LDHA-Antibody---BSA-Free-Western-Blot-NBP1-48336-img0020.jpg
Immunocytochemistry/ Immunofluorescence: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336]

Immunocytochemistry/ Immunofluorescence: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336]

Immunocytochemistry/Immunofluorescence: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336] - Immunocytochemical analysis of Lactate Dehydrogenase A in HeLa cells
Immunohistochemistry: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336]

Immunohistochemistry: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336]

Immunohistochemistry: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336] - Staining of postnatal day 8(P8) Plp-EGFP transgenic mice brain section. Image from verified customer review.
Flow Cytometry: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336]

Flow Cytometry: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336]

Flow Cytometry: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336] - An intracellular stain was performed on HeLa cells with Lactate Dehydrogenase A/LDHA Antibody NBP1-48336AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.
Western Blot: Lactate Dehydrogenase A/LDHA AntibodyBSA Free [NBP1-48336]

Western Blot: Lactate Dehydrogenase A/LDHA AntibodyBSA Free [NBP1-48336]

Western Blot: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336] - Analysis of Lactate Dehydrogenase A in HeLa whole cell lysates.
Western Blot: Lactate Dehydrogenase A/LDHA AntibodyBSA Free [NBP1-48336]

Western Blot: Lactate Dehydrogenase A/LDHA AntibodyBSA Free [NBP1-48336]

Western Blot: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336] - Analysis of Lactate Dehydrogenase A in NIH/3T3 whole cell lysates.
Western Blot: Lactate Dehydrogenase A/LDHA AntibodyBSA Free [NBP1-48336]

Western Blot: Lactate Dehydrogenase A/LDHA AntibodyBSA Free [NBP1-48336]

Western Blot: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336] - Lactate Dehydrogenase A expression in bovine skeletal muscle lysate. Image from verified customer review.
Western Blot: Lactate Dehydrogenase A/LDHA AntibodyBSA Free [NBP1-48336]

Western Blot: Lactate Dehydrogenase A/LDHA AntibodyBSA Free [NBP1-48336]

Western Blot: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336] - Lactate Dehydrogenase A expression in porcine skeletal muscle lysate. Image from verified customer review.
Immunocytochemistry/ Immunofluorescence: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336]

Immunocytochemistry/ Immunofluorescence: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336]

Lactate Dehydrogenase A-LDHA Antibody - BSA Free-Immunocytochemistry-Immunofluorescence-NBP1-48336-img0022.jpg
Immunohistochemistry: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336]

Immunohistochemistry: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336]

Immunohistochemistry: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336] - Staining of Lactate Dehydrogenase A in lung tissue.
Flow Cytometry: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336]
Simple Western: Lactate Dehydrogenase A/LDHA AntibodyBSA Free [NBP1-48336]

Simple Western: Lactate Dehydrogenase A/LDHA AntibodyBSA Free [NBP1-48336]

Simple Western: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336] - Lane view shows a specific band for Lactate dehydrogenase A in 0.1 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230kDa separation system.
Lactate Dehydrogenase A/LDHA Antibody - BSA Free

Simple Western: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336] -

Simple Western: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336] - Protein changes in ONHAs corroborate bioenergetics data. (A) Glucose transporter-1 protein levels in Stretched ONH astrocytes are significantly higher than Control (*p = 0.0225, n = 7 Control, n = 8 Stretch). Retinal lysate from a 2 month-old mouse was used as a positive control for each protein analyzed, while negative control was the signal obtained when no primary antibody was included in the capillary. (B) Lactate dehydrogenase-A, the astrocyte-specific isoform of the enzyme that catalyzes the interconversion of pyruvate & lactate, has equivalent protein levels in Control & Stretch ONH astrocytes. (C) Glucose-6-phosphate dehydrogenase, the enzyme that shunts glucose into the pentose phosphate pathway, is no different in Control & Stretch ONH astrocytes. (D) Glutamine synthetase, the enzyme that synthesizes glutamine from glutamate, is no different in Control & Stretch ONH astrocytes. (E) The monomeric form of glutamate-aspartate transporter (GLAST) has significantly higher protein levels in the Stretch as compared to the Control ONH astrocytes (p = 0.020; n = 4 Control, n = 5 Stretch). (F) GLAST dimer protein levels are no different in Control & Stretch ONH astrocytes. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35992925), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Lactate Dehydrogenase A/LDHA Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336] -

The effect of the co-culturing of cancer cells and fibroblasts on the expression of Hk2 and Ldha. (A) and (B): the co-culturing of fibroblasts (murine lung fibroblasts (MLF) or human lung fibroblasts (HLF)) with respective cancer cells (KLN205 or A549) decreased the level of Hk2 in cancer cells, having only a minor effect on the enzyme in fibroblasts. (C) and (D): in the co-cultures, the expression of Ldha was reduced in cancer cells, but was elevated in fibroblasts. Bar = 50 um. The black and white images depict differences in the shape and size of fibroblasts and cancer cells in the co-cultures. The ratio of the fluorescence of Hk2 and Ldha to beta -actin in the MLF and HLF monocultures was assumed to be 1. Asterisks indicate a statistically significant difference (p < 0.05). All the experiments were performed in triplicate and obtained similar results, and representative data from one experiment are shown in the figure. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31947613), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Lactate Dehydrogenase A/LDHA Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: Lactate Dehydrogenase A/LDHA Antibody - BSA Free [NBP1-48336] -

The effect of the co-culturing of cancer cells and fibroblasts on the expression of Hk2 and Ldha. (A) and (B): the co-culturing of fibroblasts (murine lung fibroblasts (MLF) or human lung fibroblasts (HLF)) with respective cancer cells (KLN205 or A549) decreased the level of Hk2 in cancer cells, having only a minor effect on the enzyme in fibroblasts. (C) and (D): in the co-cultures, the expression of Ldha was reduced in cancer cells, but was elevated in fibroblasts. Bar = 50 um. The black and white images depict differences in the shape and size of fibroblasts and cancer cells in the co-cultures. The ratio of the fluorescence of Hk2 and Ldha to beta -actin in the MLF and HLF monocultures was assumed to be 1. Asterisks indicate a statistically significant difference (p < 0.05). All the experiments were performed in triplicate and obtained similar results, and representative data from one experiment are shown in the figure. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31947613), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Lactate Dehydrogenase A/LDHA Antibody - BSA Free

Application
Recommended Usage

ELISA

reported in scientific literature (PMID 35052722)

Immunohistochemistry

1:100

Immunohistochemistry-Frozen

reported in scientific literature (PMID 25004202)

Immunohistochemistry-Paraffin

1:100

Simple Western

1:50

Western Blot

0.5ug/ml
Application Notes
In Western blot, a band is seen at approx. 36 kDa. 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 HeLa lysate 0.1 mg/mL, separated by Size, antibody dilution of 1:50, apparent MW was 40 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 5 reviews rated 4.4 using NBP1-48336 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

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

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: Lactate Dehydrogenase A/LDHA

Lactate dehydrogenase (LDH) is a ubiquitous enzyme that catalyzes the inter-conversion of pyruvate and lactate, with simultaneous inter-conversion of NADH and NAD+. Mammalian LDH has five tetrameric isozymes comprising combinations of two subunits (i) A subunit (LDHA or LDHM) predominates in skeletal muscle where it involve in anaerobic metabolism and pyruvate reduction (ii) B subunit (LDHB) predominates in cardiac muscle where it helps catalyze aerobic oxidation of pyruvate. Several human cancers such as including renal, breast, gastric, nasopharyngeal cancer etc. have higher LDHA levels compared to normal tissues and LDHA upregulation ensures efficient anaerobic/glycolytic metabolism for tumor cells with reduced oxygen dependency. Moreover, LDHA also plays an important role in the development, invasion and metastasis of several malignancies and induced suppression of LDHA in cancer cells has been shown to trigger ROS burst, mitochondrial pathway apoptosis and limited tumorigenic potential. Defects in LDHA have also been linked to exertional myoglobinuria and glycogen storage disease type 11 (GSD11).

Alternate Names

LDH1, LDHA, LDHM

Entrez Gene IDs

3939 (Human); 16828 (Mouse)

Gene Symbol

LDHA

UniProt

P00338 (Human)

Additional Lactate Dehydrogenase A/LDHA Products

Product Documents for Lactate Dehydrogenase A/LDHA Antibody - BSA Free

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Product Specific Notices for Lactate Dehydrogenase A/LDHA 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 Lactate Dehydrogenase A/LDHA Antibody - BSA Free

Customer Reviews for Lactate Dehydrogenase A/LDHA Antibody - BSA Free (5)

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Showing  1 - 5 of 5 reviews Showing All
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  • Lactate Dehydrogenase A/LDHA Antibody - BSA Free
    Name: Anonymous
    Application: Western Blot
    Sample Tested: glioblastoma cell lysates
    Species: Human
    Verified Customer | Posted 10/09/2023
    glioblastoma cell lysate treated with ct or drugs (A-C), 10 ug each protein were performed WB. 1/2000 dilution size markers are 37, 25, 20 kDa.
    Lactate Dehydrogenase A/LDHA Antibody - BSA Free NBP1-48336
  • Lactate Dehydrogenase A/LDHA Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HepG2 human hepatocellular carcinoma cell line and Huh-7 human hepatoma cell line
    Species: Human
    Verified Customer | Posted 03/10/2020
    Used it in 1:1000 with milk TBST.
    Lactate Dehydrogenase A/LDHA Antibody - BSA Free NBP1-48336
  • Name: Eric England
    Application: Western Blot
    Sample Tested: Whole Skeletal Muscle Lysate
    Species: Other
    Verified Customer | Posted 10/10/2014
    Lactate Dehydrogenase A/LDHA Antibody - BSA Free NBP1-48336
  • Name: Eric England
    Application: Western Blot
    Sample Tested: Whole Skeletal Muscle Lysate
    Species: Other
    Verified Customer | Posted 10/01/2014
    Lactate Dehydrogenase A/LDHA Antibody - BSA Free NBP1-48336
  • Name: Bryan Tinsley
    Application: Immunohistochemistry
    Sample Tested:
    Species: Mouse
    Verified Customer | Posted 03/17/2014
    Lactate Dehydrogenase A/LDHA Antibody - BSA Free NBP1-48336

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Protocols

View specific protocols for Lactate Dehydrogenase A/LDHA Antibody - BSA Free (NBP1-48336):

Protocol for Flow Cytometry Intracellular Staining
Sample Preparation.
1. Grow cells to 60-85% confluency. Flow cytometry requires between 2 x 105 and 1 x 106 cells for optimal performance.
2. If cells are adherent, harvest gently by washing once with staining buffer and then scraping. Avoid using trypsin as this can disrupt certain epitopes of interest. If enzymatic harvest is required, use Accutase, Collagenase, or TrypLE Express for a less damaging option.
3. Reserve 100 uL for counting, then transfer cell volume into a 50 mL conical tube and centrifuge for 8 minutes at 400 RCF.
a. Count cells using a hemocytometer and a 1:1 trypan blue exclusion stain to determine cell viability before starting the flow protocol. If cells appear blue, do not proceed.
4. Re-suspend cells to a concentration of 1 x 106 cells/mL in staining buffer (NBP2-26247).
5. Aliquot out 1 mL samples in accordance with your experimental samples.

Tip: When cell surface and intracellular staining are required in the same sample, it is advisable that the cell surface staining be performed first since the fixation and permeablization steps might reduce the availability of surface antigens.

Intracellular Staining.
Tip: When performing intracellular staining, it is important to use appropriate fixation and permeabilization reagents based upon the target and its subcellular location. Generally, our Intracellular Flow Assay Kit (NBP2-29450) is a good place to start as it contains an optimized combination of reagents for intracellular staining as well as an inhibitor of intracellular protein transport (necessary if staining secreted proteins). Certain targets may require more gentle or transient permeabilization protocols such as the commonly employed methanol or saponin-based methods.
Protocol for Cytoplasmic Targets:
Optional: Perform cell surface staining as described in the previous section.
1. Fix the cells by adding 100 uL fixation solution (such as 4% PFA) to each sample for 10-15 minutes.
2. Permeabilize cells by adding 100 uL of a permeabization buffer to every 1 x 106 cells present in the sample. Mix well and incubate at room temperature for 15 minutes.
a. For cytoplasmic targets, use a gentle permeabilization solution such as 1X PBS + 0.5% Saponin or 1X PBS + 0.5% Tween-20.
b. To maintain the permeabilized state throughout your experiment, use staining buffer + 0.1% of the permeabilization reagent (i.e. 0.1% Tween-20 or 0.1% Saponin).
3. Following the 15 minute incubation, add 2 mL of the staining buffer + 0.1% permeabilizer to each sample.
4. Centrifuge for 5 minutes at 400 RCF.
5. Discard supernatant and re-suspend in 1 mL of staining buffer + 0.1% permeabilizer.
6. Stain each sample at 1 uL/ 1 x 106 cells of primary antibody or 1-3 uL/ 1 x 106 cells for directly conjugated antibodies. Mix well and incubate at room temperature for 30 minutes- 1 hour. Gently mix samples every 10-15 minutes.
7. Following the primary/conjugate incubation, add 2 mL/sample of staining buffer +0.1% permeabilizer and centrifuge for 5 minutes at 400 RCF.
8. Remove supernatant and re-suspend each sample in 2 mL staining buffer + 0.1% permeabilizer, repeat wash for 5 minutes at 400 RCF.
9. If using a directly conjugated antibody, after the second wash, re-suspend cell pellet to a final volume of 500 uL per sample and proceed with flow analysis.

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.

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 (keep slides in the sodium citrate buffer at all times).

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


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST 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 manufacturer's instructions.

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