GAPDH Mouse mAb - Primary antibody, specific to GAPDH, IgG1

Cat. No.: Ab155825

QUALITÉ & PURETÉ Carrier Free, ExactAb™, Azide Free, Validated, High Performance, PBS Only, 1.0 mg/mL

Application

Flow Cytometry, IF/ICC, WB

Host species

Mouse

Reactive species

Human

Isotype

Mouse IgG1

Clone Number

B08/4A11

Conjugation

Unconjugated

Vue d’ensemble

Application
WB: 1/1,000-1/100,000 ; Predicted molecular weight: 37 kDa
Flow: 0.016 μg/mL-1 μg/mL
ICC/IF: 0.5 μg/mL
Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. This GAPDH antibody can be used as a loading control antibody.

Spécifications

Product Name

GAPDH Mouse mAb - Primary antibody, specific to GAPDH, IgG1

Synonymes

38 kDa BFA-dependent ADP-ribosylation substrate antibody | aging associated gene 9 protein antibody | Aging-associated gene 9 protein antibody | BARS-38 antibody | cb609 antibody | EC 1.2.1.12 antibody | Epididymis secretory sperm binding protein Li 162eP

Spécifications et pureté

Carrier Free,ExactAb™,Azide Free,Validated,High Performance,PBS Only,1.0 mg/mL

Espèce hôte

Mouse

Spécificité

GAPDH

Immunogène

Recombinant full length human GAPDH

Contrôle positif

WB: HEK293, HeLa, Jurkat, A431, HepG2 and K562 whole cell lysates; Flow: Jurkat cells; ICC/IF: HeLa cells

Conjugation

Unconjugated

Grade

Azide Free, Carrier Free, ExactAb™, High Performance, PBS Only, Validated

Isotype

IgG1

Type de chaîne lumineuse

kappa

SDS-PAGE

150 kDa

Méthode de purification

Protein A purified

Concentration

1.0 mg/mL

Product Line

Produits biologiques

Stockage et expédition

Forme

Liquid

Conditions de stockage de stockage

Store at -20°C,Avoid repeated freezing and thawing

Expédié en

Ice chest + Ice pads

Stabilité et stockage

Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.

Images

Western Blot: GAPDH Mouse mAb (Ab155825)

GAPDH Mouse mAb (Ab155825) - Western Blot
All lanes: GAPDH Antibody (Ab155825) at 0.05 μg/mL
Samples: Lysates at 20 µg per lane
Secondary: Goat Anti-Mouse IgG H&L (HRP) at 1/20000 dilution

Predicted band size: 35.8 kDa
Observed band size: 35.6 kDa
Exposure time: 36.9 s 

Flow Cytometry: GAPDH Mouse mAb (Ab155825)

GAPDH Mouse mAb (Ab155825) - Flow Cytometry
Overlay histogram showing Jurkat cells stained with GAPDH Mouse mAb (Ab155825) (Red). Cells were fixed and permeabilized with 100% methanol. A Goat Anti-Mouse IgG H&L (FITC) (Ab156254) was used as the secondary antibody. Unlabeled sample (Black) was used as a negative control.

ICC/IF: GAPDH Mouse mAb (Ab155825)

GAPDH Mouse mAb (Ab155825) - ICC/IF
IF analysis of GAPDH (Green) in HeLa cells. The cells were fixed and permeabilized with 100% methanol for 5 minutes, and blocked with 2% BSA for 1 hour at room temperature. Cells were stained with GAPDH Mouse mAb (Ab155825) at 0.3μg/mL in blocking buffer for 2 hour at RT and then incubated with Goat Anti-Mouse IgG H&L (FITC) (Ab156254) at a dilution of 1/1000 for 1 hour at room temperature in the dark. Cells were counterstained with DAPI (Blue). Images were taken on the confocal laser scanning microscope.

ICC/IF: GAPDH Mouse mAb (Ab155825)

GAPDH Mouse mAb (Ab155825) - ICC/IF
IF analysis of GAPDH (Green) in HeLa cells. The cells were fixed and permeabilized with 100% methanol for 5 minutes, and blocked with 2% BSA for 1 hour at room temperature. Cells were stained with GAPDH Mouse mAb (Ab155825) at 0.3μg/mL and Recombinant COX IV Antibody (Ab097410) (Red) at 1μg/mL in blocking buffer for 2h at room temperature and then incubated with Goat Anti-Mouse IgG H&L (FITC) (Ab156254) at a dilution of 1/1000 and Goat Anti-Rabbit IgG H&L (Cy5) (Ab170147) for 1 hour at room temperature in the dark. Cells were counterstained with DAPI (Blue). Images were taken on the confocal laser scanning microscope.

Application

Application

Dilution info

IF/ICC

0.5 μg/mL

WB

1/1000-1/100000 ; Predicted molecular weight: 37 kDa

Flow Cytometry

0.016 μg/mL- 1 μg/mL

Cibles associées (humaines)

GAPDH Tchem Glyceraldehyde-3-phosphate dehydrogenase (0 Activities)

Discover Target: GAPDH

Activity Type	Activity Value -log(M)	Mechanism of Action	Activity Reference	Publications (PubMed IDs)

Mécanismes d'action

Application Protocols

Tested applications, recommended dilutions, and protocol details are not provided in the Product Data for SKU Ab155825.

• Item-specific application data: Not specified for this item; refer to CoA/Spec Sheet.

• General guidance (literature):

  • WB: Begin titration at 1:1,000 in blocking buffer and adjust based on signal-to-noise and detection sensitivity.
  • IF/ICC: Start at 1–5 µg/mL; optimize incubation time and permeabilization conditions.
  • IHC: Optimize antigen retrieval and start with 1–10 µg/mL, adjusting for tissue and detection chemistry.
  • ELISA: Titrate in the 0.1–2 µg/mL range for capture/detection arms depending on assay design.

Always perform assay-specific optimization and include appropriate controls (isotype controls, secondary-only controls, and positive/negative biological controls).

Biological Roles

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a central enzyme in carbohydrate metabolism and exhibits multiple non-metabolic functions (general literature context).

• Primary metabolic role:

  • Catalyzes the conversion of glyceraldehyde-3-phosphate to 1,3-bisphosphoglycerate in glycolysis with concomitant reduction of NAD+ to NADH. This step links carbon flux to cellular redox state.

• Cellular localization and abundance:

  • Predominantly cytosolic; frequently abundant and relatively stable across many cell types and conditions, making it a common normalization control in immunoassays and gene expression studies (though regulation can occur under certain stresses/differentiation states).

• Moonlighting functions (literature):

  • Involvement in nuclear RNA export, DNA repair, and transcriptional regulation.
  • Participation in vesicular trafficking and cytoskeletal dynamics through protein–protein interactions.
  • Roles in redox signaling and response to oxidative stress via cysteine thiol reactivity and S-nitrosylation.

• Research implications:

  • Because GAPDH abundance can vary under specific experimental conditions (e.g., metabolic perturbation, hypoxia, differentiation), it is prudent to verify its stability before using it as a loading control in a given model system.

Note: The above pertains to the biological target (GAPDH) and provides context for selecting and interpreting results with GAPDH Mouse mAb. No therapeutic or clinical claims are made.

Buffer Applications

This product is an antibody reagent formulated in PBS and is not a buffering agent itself. Therefore, buffer preparation recipes and buffering ranges are not applicable here.

• Practical note:
  • When diluting or formulating working solutions, use compatible physiological buffers (e.g., PBS, TBS, or HEPES-buffered saline) at near-neutral pH to maintain antibody stability. Avoid introducing azide if the downstream application is azide-sensitive (e.g., HRP detection or live cells).

Green Alternatives

For antibody reagents, greener considerations focus on formulation, preservatives, shipping, and waste minimization rather than solvent substitution.

• Formulation choices:

  • Azide-free format reduces handling of acutely toxic sodium azide in the laboratory and avoids azide-containing waste streams. This product is azide-free (item-specific), which is favorable from a toxicological perspective, though aseptic technique is important to mitigate contamination risk.

• Shipping and cold chain:

  • Shipped in an ice chest with ice pads (item-specific) instead of dry ice, reducing CO2 footprint associated with dry ice sublimation and simplifying disposal.

• Concentration and aliquoting (general):

  • Concentrated, carrier-free formulations can reduce packaging and transport volume per experiment, decreasing material usage and cold-chain energy per assay.

• Comparative considerations (general):

  • Preservative: Azide-free vs azide-containing buffers.
    • Pros (azide-free): Safer waste profile; compatible with HRP and live cells.
    • Cons: Requires sterility vigilance; potentially shorter bench-life once opened.
  • Stabilizers: Carrier-free vs BSA-containing.
    • Pros (carrier-free): Lower background; amenable to conjugation; reduced animal-derived additives.
    • Cons: May require user-added stabilizers for extended storage; handle carefully to avoid aggregation.

• Waste minimization:

  • Use calibrated pipettes to avoid overuse, prepare only required working volumes, and consolidate shipments when possible to reduce packaging waste.

Pharmaceutical Uses

This product is intended strictly for research use only (from Product Data). It is not formulated, tested, or approved for clinical diagnostics, therapeutic use, or any in vivo administration.

• Laboratory/formulation context (general):
  • Carrier-free, azide-free antibodies in PBS are commonly selected for in vitro assays, conjugation to enzymes or fluorophores, and method development in research laboratories.
  • No pharmacopeial monograph or GMP status is indicated. For any regulated or translational workflows, users must perform independent qualification and consult the CoA/Spec Sheet for composition and quality attributes.

Research Use Only: Not for use in diagnostic procedures or therapeutic applications.

Physical Properties

Item-specific physicochemical specifications (appearance, concentration, excipients) are not provided in the Product Data.

• Item-specific values:

  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Formulation: PBS only (from grade note); carrier-free; azide-free.
  • Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.

• General/literature properties for mouse monoclonal IgG (for context only):

  • Typical molecular weight: ~150 kDa (IgG1/IgG2 subtypes; literature).
  • Isoelectric point (pI): commonly ~6–9 depending on isotype and sequence (literature; item-specific pI not specified).
  • Solubility: Highly soluble in aqueous buffers (PBS, TBS) with stabilizers as needed; avoid organic solvents (literature guidance).
  • Stability: Enhanced by storage at −20 °C or below; repeated freeze–thaw cycles can cause aggregation and activity loss (literature guidance).
  • Viscosity/osmolarity: buffer-dependent; not specified for this item.

• Practical notes (general):

  • Being carrier-free and azide-free, this preparation is amenable to conjugation (e.g., HRP, fluorophores) and live-cell–compatible applications that cannot tolerate sodium azide. If long-term storage at −20 °C is required, consider aliquoting to minimize freeze–thaw exposure (see Storage & Reconstitution).

Quality and Grades

This antibody is supplied with the following quality/grade attributes (from Product Data): Carrier Free; ExactAb™; Azide Free; Validated; High Performance; PBS Only.

• Carrier Free:

  • No bovine serum albumin (BSA) or other protein carriers added. Advantages: precise conjugation chemistry (e.g., NHS-ester dyes), reduced background in mass spectrometry or proteomic workflows, and minimal matrix interference.

• Azide Free:

  • No sodium azide preservative. Benefits: compatible with live-cell or enzyme-sensitive applications (e.g., HRP-based assays where azide can inhibit HRP). Tradeoff: increased need for sterile technique to prevent microbial growth.

• ExactAb™ (general description):

  • Denotes high-consistency antibody preparation optimized for lot-to-lot reproducibility and performance. Exact program specifications (isotype, clone engineering, affinity) are not specified for this item; refer to CoA/Spec Sheet.

• Validated; High Performance:

  • Indicates internal validation for performance criteria; specific assay formats, species reactivity, and data are not listed here. For detailed validation datasets, consult the product’s CoA/Spec Sheet.

• PBS Only:

  • Formulated exclusively in phosphate-buffered saline without additional stabilizers (item-specific). This simplifies downstream conjugation and assay optimization.

Recommendation: For regulated workflows, retain lot-specific CoA/Spec Sheet and perform in-house qualification under your assay conditions.

Reaction and Applications

As a research antibody, this product is intended for immunodetection and capture workflows rather than chemical synthesis.

• Research applications (general/literature for anti-GAPDH mAbs):

  • Western blot (WB): GAPDH serves as a cytosolic housekeeping/loading control in many mammalian lysates due to abundant, relatively constant expression.
  • Immunofluorescence (IF) and Immunocytochemistry (ICC): Cytoplasmic staining patterns in many cell lines and primary cells; useful for normalization or localization studies.
  • Immunohistochemistry (IHC): Detection in paraffin-embedded or frozen tissues after appropriate antigen retrieval (clone- and protocol-dependent).
  • ELISA/Capture: May be employed in sandwich or indirect ELISA formats for GAPDH quantitation in research samples.
  • Flow cytometry: Less common but feasible for permeabilized cells to measure intracellular GAPDH.

• Practical tips (general):

  • Optimize antibody dilution empirically based on sample load and detection system. Titrate to achieve linear signal for normalization.
  • Validate lysis buffer and loading amounts to avoid signal saturation with abundant GAPDH.
  • Use appropriate secondary antibodies matched to the mouse isotype (isotype for this item is not specified; verify on CoA/Spec Sheet).

Note: Specific, tested applications for SKU Ab155825 are not listed in the Product Data; consult the CoA/Spec Sheet for item-validated applications and recommended dilutions.

Reaction Conditions

Classical chemical reaction conditions (solvent, temperature, catalysts) are not applicable to antibody usage. However, practical assay conditions for immunoapplications are noted below as general guidance (literature-based; item-specific parameters are not provided in the Product Data):

• Western blot (general):

  • Sample: whole-cell or tissue lysates (10–40 µg total protein per lane commonly used). Denaturing SDS-PAGE, transfer to PVDF/nitrocellulose. Block with 3–5% milk or BSA. Primary incubation typically 1–16 h at 4–25 °C. Secondary HRP- or fluorophore-labeled anti-mouse IgG.

• Immunofluorescence/ICC (general):

  • Fixation: 4% paraformaldehyde or methanol; permeabilization with 0.1–0.3% Triton X-100 as needed. Block in serum/BSA. Incubate primary 1–2 h RT or overnight 4 °C.

• IHC (general):

  • Antigen retrieval: heat-induced epitope retrieval in citrate or EDTA buffers; conditions must be optimized. Detection via polymer-based HRP or fluorescence systems.

• ELISA (general):

  • Coating: 0.5–2 µg/mL capture; detection antibody and standards titrated empirically.

Item-specific, validated conditions, working dilutions, and species reactivity for SKU Ab155825 are not specified; consult the CoA/Spec Sheet and perform in-house optimization.

Safety and Handling

Safety data specific to this item are not provided in the Product Data and should be confirmed in the SDS.

• GHS classification and hazard statements (item-specific):

  • Signal word: Not specified for this item; refer to SDS.
  • H-statements: Not specified for this item; refer to SDS.
  • GHS classification: Not specified for this item; refer to SDS.
  • Pictograms: Not specified for this item; refer to SDS.

• General laboratory safety guidance for antibody solutions:

  • PPE: Laboratory coat, gloves, and safety glasses are recommended when handling biological protein preparations and buffers.
  • Avoid ingestion, inhalation of aerosols, and contact with skin/eyes. Wash thoroughly after handling.
  • Spill response: Absorb with inert material, clean with water/detergent. Dispose according to institutional biosafety procedures.
  • Incompatibilities: Avoid strong oxidizers and extremes of pH or temperature that denature proteins. Avoid microbial contamination (azide-free formulation lacks a common preservative).
  • Special risks: Repeated freeze–thaw can cause aggregation and loss of binding activity. Use aseptic technique to maintain integrity during aliquoting and use.

• First aid (general):

  • Skin/eye contact: Rinse with water for several minutes; seek medical attention if irritation persists.
  • Ingestion/inhalation: Move to fresh air; seek medical advice if unwell.

Always consult the SDS for authoritative, item-specific safety information.

Solvent Selection

This product is an aqueous antibody preparation formulated in PBS only (from Product Data). Organic solvent selection is not applicable.

• Aqueous buffer compatibility (general guidance):

  • Compatible: PBS, TBS, HEPES-buffered saline for dilution. Maintain near-neutral pH to preserve antibody integrity.
  • Avoid: Strongly acidic/basic solutions, chaotropes, and high percentages of organic solvents that can denature or precipitate IgG.

• Additives (general, if needed by the user):

  • Since the product is carrier-free and azide-free, users may optionally add qualified stabilizers post-aliquoting (e.g., 0.1–1% sterile-filtered BSA or 10–50% glycerol) depending on application. Such modifications should be validated in-house.

• When to choose this format:

  • Choose PBS-only, carrier-free, azide-free formulations when planning covalent conjugation (e.g., fluorophores, enzymes), when azide is incompatible (e.g., HRP-based detection), or when minimizing exogenous proteins is required (e.g., proteomics).

• Comparison (general):

  • Carrier-free vs. carrier-containing: Carrier-free reduces background and enables conjugation; carrier-containing may enhance stability for routine storage but can interfere with labeling or certain assays.

Storage and Reconstitution

Item-specific storage conditions (from Product Data):

• Storage: Store at −20 °C; avoid repeated freezing and thawing.
• Shipment: Ice chest + ice pads.
• Formulation cues: PBS only; carrier-free; azide-free. Exact concentration/appearance not specified; refer to CoA/Spec Sheet.

Handling and stability guidance (general best practices for antibodies):

• Upon receipt: Inspect for container integrity. If the product is liquid, briefly centrifuge to collect contents.
• Aliquoting: Divide into single-use aliquots under sterile conditions to prevent contamination and to avoid freeze–thaw cycles that can reduce binding activity.
• Thawing: Thaw on ice; gently mix by inversion. Do not vortex vigorously.
• Working solutions: Prepare dilutions in cold PBS or assay buffer immediately before use. Discard unused working dilutions; do not return to stock vial.
• Optional user-added stabilizers (if compatible with application): 10–50% glycerol for storage at −20 °C, or 0.1–1% BSA to reduce adsorption. Validate impacts on downstream assays before routine use.
• Light and temperature: Protect from prolonged ambient temperatures and light exposure, especially if conjugated post-purchase.

For exact concentration, isotype, and any reconstitution steps (if lyophilized), consult the lot-specific CoA/Spec Sheet.

Structure and Identity

Monoclonal antibody reagent targeting glyceraldehyde-3-phosphate dehydrogenase (GAPDH), supplied as an azide-free, carrier-free preparation in PBS (ExactAb™, Validated, High Performance).

• Item-specific facts (from Product Data):

  • Product: GAPDH Mouse mAb (SKU: Ab155825)
  • Host/Type: Mouse monoclonal antibody (mAb)
  • Grade/Purity: Carrier Free; ExactAb™; Azide Free; Validated; High Performance; PBS Only
  • CAS, CID, SMILES, InChIKey: Not applicable for protein biologics

• Structural features (general antibody knowledge):

  • Immunoglobulin G (IgG)-type architecture (literature): two identical heavy chains (~50 kDa each) and two identical light chains (~25 kDa each) forming a Y-shaped heterotetramer with interchain disulfide bonds.
  • Antigen-binding regions (Fab) contain variable domains (VH/VL) conferring specificity to GAPDH epitopes; Fc mediates secondary detection and purification interactions (e.g., Protein A/G binding; isotype-dependent, not specified for this item).

• Molecular metrics (literature/typical):

  • Approximate MW of a typical mouse IgG: ~150 kDa (literature). Exact mass/isotype for this specific clone: Not specified for this item; refer to CoA/Spec Sheet.

• Antigen context (general knowledge):

  • GAPDH is a cytosolic enzyme involved in glycolysis and is abundantly expressed in many cell types, often used as a loading control in immunoassays. Specific epitope/clone information is not provided for this item.

Synthetic Utility

Not applicable. This product is a monoclonal antibody intended for immunodetection and capture assays, not for chemical synthesis or as a small-molecule building block.

• For relevant information, see:
  • Reaction & Applications (immunoassays and detection)
  • Application Protocols (if provided on CoA/Spec Sheet)
  • Storage & Reconstitution (handling guidance)

Target Specificity

Only item-specific details from the Product Data can be listed here.

• Antigen/Target: GAPDH (glyceraldehyde-3-phosphate dehydrogenase)
• Host/Type: Mouse monoclonal antibody (mAb)
• Clone, epitope, isotype, species reactivity, cross-reactivity, and binding affinity: Not specified for this item; refer to CoA/Spec Sheet.

Note: Without clone and isotype information, users should verify secondary antibody compatibility (anti-mouse IgG isotype selection) and confirm species reactivity in pilot experiments.

Foire aux questions

What does ExactAb? mean?

ExactAb? indicates the product has been validated for use in rigorous antibody validation workflows. Validation may include positive/negative control samples, signal-to-noise testing, and protocol-specific QC. Lot-specific test data is published on the Certificate of Analysis where available.

What does Carrier Free mean?

Carrier Free indicates that the material has been processed and tested for specialty industrial processing. Specifications focus on the impurities that affect performance in that application — for example, metal content for electronics, particle size for batteries, refractive index for optics, or moisture content for moisture-sensitive processes.

How should this product be stored?

Store at ?20 °C. Avoid repeated freeze–thaw cycles. Aliquot into single-use volumes before freezing.

How is this product shipped?

This product ships in an insulated container with ice pads. Unpack on arrival and transfer it to the storage condition stated above.

What documentation is provided?

Available product documentation, including Certificates of Analysis (COA), Safety Data Sheets (SDS), and specification sheets, is shown in the product document area. Document availability and access follow the current site policy.

Which species reactivity has this antibody been validated against?

Validated reactivity is listed on the PDP and on each lot's datasheet. We validate against the most commonly tested species for the target (GAPDH Mouse mAb); cross-reactivity in untested species may exist but is not guaranteed. Contact technical support for specific species inquiries.