Guinea Pig IgG - Isotyp-Kontrollantikörper, IgG

Cat. No.: Ab175978

GRADE & PURITY Carrier Free, ExactAb™, Azide Free, Validated, PBS Only, ≥95%, See COA

Application

CLIA, EIA, LF

Host species

Meerschweinchen

Isotype

Guinea pig IgG

Conjugation

Unconjugated

Übersicht

Guinea Pig IgG Antibody; Isotype Control Antibody

Specifications

Product Name

Guinea Pig IgG - Isotyp-Kontrollantikörper, IgG

Synonyme

Guinea Pig IgG

Spezifikationen & Reinheit

Carrier Free,ExactAb™,Azide Free,Validated,PBS Only,≥95%,See COA

Wirtsspezies

Meerschweinchen

Konjugation

Unconjugated

Note

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

Isotyp

IgG

SDS-PAGE

150 kDa

Verfahren zur Reinigung

Affinity chromatography

Konzentration

See COA

Product Line

Biologische Produkte

Lagerung und Versand

Form

Liquid

Storage

Store at -20°C

Verschickt in

Ice chest + Ice pads

Stabilität und Lagerung

Store at 4°C short term (1-2 weeks). Store at -20°C long term (24 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.

Bilder

SDS-PAGE: Guinea Pig IgG (Ab175978)

Guinea Pig IgG (Ab175978) - SDS-PAGE
3 μg/lane of Guinea Pig IgG (Ab175978) was resolved with SDS-PAGE under reducing (R) and non-reducing (N) conditions and visualized by Coomassie® Blue staining. Showing the band at 26.1 kDa (Light Chain) & 51.3 kDa (Heavy Chain), under reducing conditions; 161.4 kDa, under non-reducing conditions. The purity of the Guinea Pig IgG is greater than 95%.

Application Protocols

• Item-specific tested applications, recommended dilutions, and positive controls: Not specified for this item; refer to CoA/Spec Sheet.
• General usage guidance (literature/practice; tailor to your assay)
  • ELISA standard/control: prepare serial dilutions in sample diluent (PBS + 1% BSA). Coat or capture as per platform; detect with anti–guinea pig IgG secondary. Optimize coating concentrations (0.5–2 µg/mL) and detection ranges empirically.
  • Western blot blocking/control: use as species control to evaluate secondary specificity. Block membranes with non-interfering blockers; probe with anti–guinea pig secondary to assess background.
  • Immunofluorescence/flow cytometry: include a guinea pig IgG isotype control at the same concentration as the test primary to gauge non-specific signals; detect with cross-adsorbed anti–guinea pig secondary.
  • Conjugation: see Reaction Conditions for typical labeling parameters; confirm DOL and retain activity.
• Validation note
  • Because “Validated” is indicated without specifics, confirm performance in your exact matrix and detection system and document lot-specific conditions for reproducibility.

Biological Roles

• Overview (literature/general)
  • Immunoglobulin G (IgG) is the predominant antibody class in mammalian serum, mediating humoral immunity. It neutralizes pathogens, opsonizes antigens for phagocytosis, and can activate complement via the classical pathway.
  • Structure–function: variable regions (Fab) confer antigen specificity through somatic recombination and hypermutation; the Fc region engages Fcγ receptors and complement component C1q, orchestrating effector responses.
  • Guinea pig IgG shares conserved immunoglobulin architecture with species-specific sequence and glycosylation patterns that modulate Fc receptor interactions and Protein A/G binding.
• Biochemical features (literature)
  • N-linked glycosylation at Fc CH2 domain influences stability and effector function.
  • pH-dependent Fc receptor interactions underpin endosomal recycling pathways in some species (FcRn), affecting half-life of IgG.
• Relevance in research
  • Serves as a species control, standard, or immunogen; provides a platform for studying cross-species Fc interactions, complement activation, and antibody engineering.
• Important caveats
  • Biological function and in vivo effects depend on subclass, glycoform, and context; these attributes are not specified for this item and should not be inferred without data.

Buffer Applications

• Item-specific note: Product is indicated as “PBS Only.” Exact pH and ionic strength are not specified; refer to CoA/Spec Sheet.
• General guidance (literature/practice)
  • Phosphate-buffered saline (PBS) at pH ~7.2–7.4 stabilizes IgG near physiological conditions and supports binding in most immunoassays.
  • For ELISA and plate-based assays:
    • Coating: carbonate–bicarbonate buffer, pH 9.4–9.6, can improve passive adsorption of capture antibodies (not item-specified; prepare by user if needed). After coating, switch to PBS or TBS with 0.05% Tween-20 for washing.
    • Blocking: protein-based blockers (BSA, casein) or synthetic blockers reduce nonspecific binding. Avoid carriers if maintaining carrier-free conditions is critical.
  • For conjugation: exchange to application-appropriate buffers (e.g., PBS→amine-free bicarbonate for NHS ester reactions; PBS→HEPES for metal-sensitive steps). Remove azide-sensitive contaminants if present in user-added reagents.
  • For storage: some workflows add 2–5% trehalose or 10–50% glycerol at −20°C to limit freeze–thaw damage (glycerol not advisable if maintaining carrier/glycerol-free status is required by the application).

Green Alternatives

• Product-intrinsic considerations
  • Azide Free: eliminates sodium azide, a toxic preservative, improving environmental and safety profiles in downstream waste streams and enabling enzyme-based detections without azide quenching.
  • Carrier Free: reduces proteinaceous additives, simplifying waste and minimizing interference—often permitting lower reagent consumption through improved signal-to-noise.
• Greener handling practices (general)
  • Use reusable, low-binding containers where appropriate and compatible with sterility.
  • Favor aqueous buffer exchange over precipitation with organic solvents.
  • Optimize assay designs to minimize antibody consumption (microfluidics, high-binding plates, miniaturized volumes).
  • Shipments already use cold packs; consolidate orders to reduce packaging impact when feasible.
• Comparison (general, literature)
  • Azide-preserved antibodies vs azide-free
    • Pros of azide-free: compatible with HRP/AP conjugates, live cells, copper-catalyzed click pre-reactions; reduced hazardous preservative waste.
    • Cons: shorter open-vial stability; requires stricter aseptic handling.
  • Carrier-containing vs carrier-free
    • Pros of carrier-free: cleaner conjugation, lower background in MS/SEC; easier quantification.
    • Cons: greater adsorption losses; may benefit from low concentrations of inert protein or trehalose during storage (user-added, if assay-compatible).
• Net assessment
  • For most immunochemical workflows, azide-free, carrier-free IgG is the more sustainable and versatile choice when paired with careful aliquoting and cold-chain stewardship.

Pharmaceutical Uses

• Scope limitation: This product is for research use only. No clinical or therapeutic use is claimed or supported.
• Relevant non-clinical/industrial contexts (general)
  • Assay reagent: used to develop and qualify analytical methods (ELISA, SPR/BLI) as a reference IgG material for platform performance, specificity testing, and matrix effect assessments.
  • Process development: model protein for purification platform studies (Protein A/G capture, ion exchange optimization, viral clearance validation) in bioprocess R&D.
  • Diagnostic development (research-phase): control antibody for immunoassay calibration, stability studies, and lot-to-lot comparability testing.
  • Conjugate manufacturing (research): starting material for producing labeled secondaries or tracers (biotinylated, enzyme- or fluorophore-conjugated) for kit prototyping.
• Regulatory note
  • No pharmacopeial monograph or GMP status is provided. Any use in regulated workflows requires independent qualification. Lot-specific attributes (purity, identity, bioburden, endotoxin) are Not specified for this item; refer to CoA/Spec Sheet if required.

Physical Properties

• Item-specific properties
  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular weight (exact): Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
  • Buffer: “PBS Only” indicated; detailed composition (pH, ionic strength) not specified for this item; refer to CoA/Spec Sheet.
• Literature/general characteristics for IgG proteins (for experimental planning; not item specifications)
  • Typical molecular mass: ~150 kDa (∼50 kDa heavy chains, ∼25 kDa light chains under reducing SDS-PAGE).
  • Isoelectric point (pI): typically ~6–9 depending on subclass and glycoform.
  • Solubility: highly soluble in isotonic aqueous buffers (e.g., PBS, ~1–10 mg/mL and higher), salting-out in high ammonium sulfate.
  • Stability range: best maintained near neutral pH (6.8–7.6) with moderate ionic strength; susceptible to aggregation upon freeze–thaw, agitation, or exposure to interfaces.
  • Extinction coefficient (approximate, human IgG1 reference): ε280 ≈ 203,000 M−1 cm−1; A280 of 1.0 ≈ 0.73 mg/mL (literature, sequence-dependent). Not a specification for this item.
  • Temperature sensitivity: retains structure at 2–8°C short-term; long-term storage typically −20°C to −80°C with cryoprotectants (literature).
  • Denaturants: unfolded by chaotropes (urea, guanidinium), reduced by DTT/TCEP, digested by proteases (papain, pepsin) in a controlled manner (literature).

Quality and Grades

• Item-specific grade descriptors (from Product Data)
  • Carrier Free: supplied without protein carriers (e.g., BSA, gelatin) and typically without glycerol. Advantageous for conjugation (biotin, fluorophores, enzymes) and mass-sensitive assays where carriers interfere. Also reduces non-specific background from carrier proteins.
  • ExactAb™: branded designation by the supplier; indicates a defined, quality-controlled antibody format. Exact technical attributes for this lot are not specified here; refer to CoA/Spec Sheet for details.
  • Azide Free: no sodium azide preservative, enabling compatibility with peroxidase-based detections, live-cell exposures, and conjugation chemistries sensitive to azide. Requires aseptic technique and proper storage to maintain sterility/stability.
  • Validated: indicates performance verification by the supplier. Specific validated applications and dilutions are not provided here; refer to CoA/Spec Sheet.
  • PBS Only: formulated in phosphate-buffered saline. Exact pH, salt concentration, and any stabilizers are not specified here; refer to CoA/Spec Sheet.
• What these grades mean in practice
  • Conjugation-ready: absence of carriers/azide minimizes side reactions during NHS-ester, maleimide, click, or enzymatic labeling.
  • Analytical use: low excipient background helps in ELISA, SPR, BLI, HPLC-SEC, and MS workflows.
  • Handling implication: azide-free, carrier-free antibodies demand careful aliquoting and sterile technique to prevent contamination and proteolysis.
• Lot-specific controls
  • Concentration, purity by SDS-PAGE/SEC, endotoxin, and bioburden: Not specified for this item; refer to CoA/Spec Sheet.

Reaction and Applications

• This section focuses on experimental applications of Guinea Pig IgG as a research reagent (general literature guidance). No specific validated uses are claimed for this item beyond the catalog descriptors.
• Common applications
  • Immunoassays: ELISA/ELISPOT plate blocking controls, isotype-matched control in flow cytometry or immunofluorescence (when a guinea pig primary is used), and calibration standards.
  • Affinity workflows: immunogen for raising anti–guinea pig secondary antibodies in other host species; ligand in Fc-binding studies (Protein A/G/L binding, species-dependent).
  • Conjugate development: starting material for labeling with fluorophores (e.g., Alexa Fluor dyes), biotin, enzymes (HRP, AP), or haptens for detection or pull-down assays.
  • Structural/biophysical: SEC-MALS and DSC calibration, SPR/BLI method development using a well-behaved IgG.
• Practical tips (literature)
  • Minimize freeze–thaw; aliquot upon receipt. Mix gently—avoid vortexing to limit aggregation.
  • For NHS-ester labeling, exchange into amine-free buffer (e.g., 50 mM sodium bicarbonate, pH ~8.3). For thiol-directed labeling, generate free thiols on hinge (mild reduction) and use maleimides; remove excess reagents by desalting.
  • Validate Fc-binding with Protein A/G empirically, as guinea pig IgG shows species-specific binding profiles.
  • Use matched secondary antibodies (anti–guinea pig IgG) to avoid cross-species cross-reactivity in multiplexed assays.

Reaction Conditions

• Note: No item-specific validated protocols are provided. The following are general literature conditions for common IgG manipulations.
• Biotinylation (NHS-ester)
  • Buffer: 50 mM sodium bicarbonate, pH 8.3; IgG 0.5–2 mg/mL. Molar ratio dye:IgG 3–10:1. React 30–60 min at room temperature. Quench with Tris or glycine; desalt.
• Fluorophore/enzyme conjugation (maleimide)
  • Generate thiols: reduce hinge with 1–5 mM TCEP for 30–60 min at RT; remove reductant. React with maleimide dye at pH 6.5–7.0 for 1–2 h. Purify by SEC.
• Immobilization on beads/plates
  • Passive adsorption: carbonate buffer pH 9.6, 0.5–5 µg/mL coating overnight at 4°C. Block and wash in PBS/TBS + 0.05% Tween-20.
  • Covalent coupling: NHS-activated resin; couple in 0.1 M MES, pH 5.0–6.0 for 1–2 h; cap and wash.
• Protein A/G capture (species-dependent)
  • Equilibrate resin in PBS; load IgG at 1–10 mg/mL; wash; elute with glycine pH 2.7–3.0 for 1–3 min; neutralize immediately. Confirm compatibility with guinea pig IgG binding empirically.
• Storage during procedures
  • Keep on ice or at 2–8°C; avoid vigorous mixing. Add 2–5% trehalose for freeze–thaw protection when storing at −20°C (if compatible with “carrier-free” requirements).

Safety and Handling

• GHS classification and hazard statements for this item: Not specified for this item; refer to SDS.
• Signal word, pictograms, H-statements: Not specified for this item; refer to SDS.
• General safety guidance for protein antibodies (literature/practice)
  • Although azide-free (per Product Data), handle as a laboratory protein reagent. Avoid ingestion, inhalation of aerosols, and contact with eyes/skin. Follow institutional biosafety procedures.
  • PPE: lab coat, gloves (nitrile), and safety glasses. Use biological safety cabinet if aerosol generation is possible.
  • Incompatibilities: avoid strong acids/bases (can denature protein), oxidizing agents, proteases, and organic solvents that precipitate proteins. Do not mix with sodium azide if downstream applications are azide-sensitive (e.g., HRP-based detection) or for live-cell exposure.
  • First aid (general): rinse affected skin/eyes with water; if inhaled, move to fresh air; if ingested, rinse mouth. Seek medical attention as needed. Defer to SDS for authoritative instructions.
  • Spill response: absorb with inert material, clean with aqueous buffer/detergent, disinfect surfaces if biological contamination is a concern.
  • Special risks: repeated freeze–thaw can generate particulates/aggregates; minimize to prevent unexpected immunogenicity or assay background.
  • Waste: dispose of aqueous protein waste per local regulations; decontaminate containers before disposal.

Solvent Selection

• Applicability note: This product is an aqueous protein reagent; organic solvent selection is generally not applicable. Focus on buffer compatibility and mild additives.
• Buffer system
  • Supplied in PBS (item-specific). Typical PBS is ~10 mM phosphate, 137 mM NaCl, pH 7.2–7.4 (literature; exact composition for this item not specified).
  • Compatible with low levels of non-ionic surfactants (e.g., 0.01–0.05% Tween-20) to reduce adsorption, if compatible with downstream assays.
• Avoid
  • Strong organic solvents (acetone, acetonitrile, methanol) that precipitate/denature IgG.
  • Extreme pH (<5 or >9) and high ionic strength unless intentionally used for elution in affinity workflows.
• When to adjust buffer
  • Conjugation: exchange into amine-free (for NHS-ester) or reducing-agent-free (for maleimide) buffers using spin desalting or dialysis.
  • Cell-based assays: maintain isotonicity and physiological pH. Consider adding stabilizers such as 2–5% trehalose for freeze–thaw protection (literature guidance).
• Quick comparison (general)
  • PBS vs TBS: TBS can reduce phosphate-related background in alkaline phosphatase assays; PBS is preferred for many mammalian systems. Choose based on enzyme conjugate and assay chemistry.

Storage and Reconstitution

• Item-specific storage and shipping
  • Storage conditions: Store at −20°C (per Product Data). Avoid repeated freeze–thaw cycles.
  • Shipped in: Ice chest + Ice pads (per Product Data). Inspect upon receipt to ensure cold-chain integrity.
• Formulation details
  • Grade: Carrier Free, Azide Free, PBS Only (per Product Data). Exact concentration, pH, and any stabilizers: Not specified for this item; refer to CoA/Spec Sheet.
• Handling recommendations (general best practice)
  • Upon first thaw, gently mix by inversion; avoid vortexing/foaming. Centrifuge briefly to collect contents.
  • Aliquot into single-use volumes under sterile conditions to minimize freeze–thaw and contamination. Use low-binding tubes to reduce adsorption losses (especially for carrier-free formulations).
  • Short-term storage: 2–8°C for hours to a few days, as assay permits.
  • Long-term storage: −20°C; for multi-month storage, consider −80°C. If compatible with your application, user-added stabilizers such as 2–5% trehalose can improve freeze–thaw resilience (literature).
  • Do not store with sodium azide if subsequent HRP-based detection or live-cell exposure is planned.
• Reconstitution
  • If provided lyophilized (not specified here), reconstitute in sterile PBS or assay buffer to the desired concentration with gentle mixing. Since the state (liquid/lyo) is not specified for this item, refer to CoA/Spec Sheet for exact instructions.
• Documentation
  • Record aliquot concentrations and storage dates; verify integrity by SDS-PAGE or functional assay before critical experiments.

Structure and Identity

• Item-specific identifiers
  • Product name: Guinea Pig IgG (polyclonal immunoglobulin G)
  • SKU: Ab175978
  • Category: Antibody reagent (research use only)
  • CAS, CID, InChIKey, SMILES: Not specified for this item; refer to CoA/Spec Sheet.
• Composition and architecture (general, literature)
  • Immunoglobulin G is a heterotetrameric glycoprotein (~150 kDa, literature) composed of two identical heavy (γ) chains and two identical light (κ or λ) chains linked by disulfide bonds.
  • Modular domains: two antigen-binding Fab arms (variable heavy/light domains) and one Fc region (constant domains) mediating effector functions.
  • Post-translational features: conserved N-glycosylation in Fc (species- and subclass-dependent; literature).
  • Quaternary structure is Y-shaped in 2D depictions: hinge connects Fab arms to Fc; disulfides stabilize the assembly.
• Species/source context (general)
  • “Guinea Pig IgG” refers to total/purified IgG from guinea pig, typically used as an immunochemical reagent, control, or immunogen (literature). Subclass distribution (e.g., IgG1/IgG2) and light chain type are species-dependent and not item-specified here.
• Molecular formula and exact molecular weight for this specific lot: Not specified for this item; refer to CoA/Spec Sheet.
• Structural features relevant to handling (general)
  • Proteinaceous, amphiphilic macromolecule; sensitive to denaturation by extremes of pH, temperature, organic solvents, and surfactants at high concentrations (literature).

Synthetic Utility

• Applicability: For antibodies, “synthetic utility” maps to bioconjugation and protein engineering rather than small-molecule synthesis.
• Conjugation chemistries (general, literature)
  • Lysine-directed: NHS-ester activated dyes/biotin react with surface amines at pH 8.3–8.5; broad labeling with potential impact on antigen binding.
  • Cysteine-directed: maleimide/iodoacetamide reagents couple to reduced hinge thiols; offers site-enriched labeling after partial reduction.
  • Carbohydrate-directed: Fc glycan oxidation (NaIO4) followed by oxime/hydrazone ligation enables Fc-selective conjugation.
  • Enzymatic: sortase A, microbial transglutaminase, and glycan remodeling (EndoS2) provide higher site-specificity.
• Downstream utilities
  • Creation of secondary reagents (anti–guinea pig detection probes), imaging agents, pull-down/affinity resins (via biotin–streptavidin or direct immobilization), and assay controls.
• Practical considerations
  • Buffer exchange to remove amines/reducing agents before conjugation; control degree of labeling (DOL) spectrophotometrically.
  • Verify function post-labeling (antigen binding, if applicable; Fc interactions) and remove aggregates by SEC.
  • For immobilization, use oriented capture via Protein A/G when compatible with guinea pig Fc binding; otherwise use random covalent coupling (e.g., epoxy or NHS-activated surfaces).

Target Specificity

• Item-specific target information: Not specified for this item; refer to CoA/Spec Sheet.
• Context (general)
  • The product name “Guinea Pig IgG” indicates a purified immunoglobulin from guinea pig, not an antibody raised against a defined antigen. Therefore, no single antigen specificity, clone, or isotype subclass is implied here.
  • When used as a control, match species and isotype to the experimental primary to assess non-specific binding of secondary antibodies or Fc-mediated interactions.
  • For detection, pair with anti–guinea pig secondary antibodies validated in your assay format (WB, IHC, IF, FC).