HSP25/HSP27 Antibody

What it means: Target is the antigen this antibody is intended to recognize. The target on this page is HSP25/HSP27.

How to interpret here: Use HSP25/HSP27 as your primary “match checkpoint.” Confirm the symbol/name aligns with the protein you need to detect, and be cautious with closely related family members or isoforms that can share epitopes. Immunogen is Human HSP27 peptide (species: Human). Immunogen context helps you anticipate epitope coverage and potential cross-reactivity (e.g., conserved regions vs unique segments). Clonality is Monoclonal (clone 8A7), which supports long-term reproducibility when reordering and comparing across studies. Reactivity is listed as Human, Mouse, Rat, Bovine, Dog, Guinea Pig, Hamster, Sheep. Choose a species match for your samples; treat unlisted species as requiring independent validation. Host is Mouse—this guides secondary antibody selection and multiplex planning. Isotype is IgG1 Kappa; match this when choosing subclass-specific secondaries or isotype controls. Conjugate options include Unconjugated; ATTO 390; ATTO 488; ATTO 594; APC; Biotin; FITC; HRP; PCP; RPE. Conjugation changes detection strategy (direct vs secondary-based) and impacts panel design. Listed applications are WB, IHC, ICC/IF, IP, FCM, FACS. Use these as a validation scope for your workflow and sample type. Supplier notes: Detects ~25kDa or ~27kDa. Recognizes HSP25 and HSP27, cross reacts with alpha B crystallin.. Use these details to interpret bands/staining patterns and plan controls. Purification is Protein G Purified—purification level can influence background in sensitive assays. Concentration is 1 mg/ml; treat this as formulation information and still titrate in your assay for best signal-to-background.

Why it matters for buying:

• Specificity management: Target identity and immunogen context reduce the risk of cross-reactivity and misleading bands/staining.
• Species fit: Reactivity determines whether binding is likely in your organism and matrix.
• Workflow fit: Validation in your intended applications reduces optimization burden and failed experiments.
• Reproducibility: Clone number (for monoclonals) helps you keep results comparable over time and across labs.

Practical guidance:

• Confirm the biological form: If your study depends on a specific isoform/PTM or processed form, verify the antibody’s epitope context (immunogen/notes) aligns with that form.
• Plan specificity controls: Use known positive/negative samples and, when feasible, KO/KD/overexpression controls to confirm target-specific signal.
• Titrate for your matrix: Antibody performance is condition-dependent—optimize dilution to reduce background while preserving signal.
• Choose detection format intentionally: Use unconjugated + secondary for flexibility/amplification, or direct conjugates for faster workflows and multiplexing.

What to watch out for:

• Family-member cross-reactivity: Closely related proteins can yield off-target bands or staining; interpret multi-band patterns with controls.
• Fixation/denaturation effects: Epitopes can be masked or altered across WB vs IF/IHC; validate under your exact prep conditions.
• Cross-species assumptions: Do not assume reactivity in unlisted species without testing.

What it means: Clone number identifies the specific monoclonal antibody clone. The clone listed here is 8A7.

How to interpret here: Use 8A7 as the identifier for reproducibility—clone names help you match literature, compare vendors, and reorder the same binding behavior over time. Isotype is IgG1 Kappa; this matters for subclass-specific secondaries, isotype controls, and Fc-binding reagents (Protein A/G). Conjugate options include Unconjugated; ATTO 390; ATTO 488; ATTO 594; APC; Biotin; FITC; HRP; PCP; RPE. The same clone may be available in different conjugated formats; detection changes but epitope binding is typically the same. Applications listed: WB, IHC, ICC/IF, IP, FCM, FACS. Confirm this clone is supported for your intended method and sample prep. Supplier notes: Detects ~25kDa or ~27kDa. Recognizes HSP25 and HSP27, cross reacts with alpha B crystallin.—use these to interpret expected band size or specificity caveats.

Why it matters for buying:

• Reproducibility: Clone identity is the most reliable way to obtain the same monoclonal specificity.
• Panel design: In multiplex staining/flow, clone choice affects epitope competition and staining intensity.
• Continuity: Labs often standardize on clone names; matching clone reduces retraining and re-optimization.

Practical guidance:

• Document the clone: Record clone + lot + dilution in your methods to preserve reproducibility.
• Check epitope competition: For multi-antibody panels, avoid using two clones that compete for the same epitope on the same target.
• Choose conjugate to match platform: For direct detection, pick a conjugate compatible with your instrument/filters and compensation plan.

What to watch out for:

• Same target, different clones: Antibodies to the same target are not interchangeable—clone behavior can differ by application and sample prep.
• Lot effects: Monoclonals are generally consistent, but conjugation and handling can still shift performance—revalidate when switching lots for quantitative work.

What it means: Clonality describes whether the antibody comes from a single clone (monoclonal) or a mixture recognizing multiple epitopes (polyclonal). The value shown here is Monoclonal.

How to interpret here: Use Monoclonal to choose between epitope precision vs broader epitope coverage. Clonality can affect specificity, robustness across sample prep, and lot-to-lot consistency. This is a monoclonal with clone 8A7, which supports reproducibility and consistent epitope recognition. Monoclonals often provide cleaner specificity and are easier to standardize for quantitative comparisons. Applications listed: WB, IHC, ICC/IF, IP, FCM, FACS. If your epitope is sensitive to fixation/denaturation, polyclonals sometimes perform better; for standardized quantitation, monoclonals are often preferred. Host is Mouse—plan secondary antibodies accordingly. Reactivity is Human, Mouse, Rat, Bovine, Dog, Guinea Pig, Hamster, Sheep; clonality does not guarantee cross-species performance—validate per species.

Why it matters for buying:

• Specificity vs sensitivity: Monoclonals favor specificity; polyclonals may increase sensitivity/robustness.
• Reproducibility: Clone-defined reagents simplify long-term consistency and cross-lab comparisons.
• Assay dependence: Some assays tolerate multi-epitope binding better than others (e.g., IHC/IF vs quantitative WB).

Practical guidance:

• If you need consistency: Prefer monoclonal + clone number, and record lot information for publications.
• If signal is difficult: Consider whether multi-epitope recognition (polyclonal) could help under your prep conditions.
• Validate with controls: Use KO/KD, peptide blocking (when appropriate), or orthogonal methods to support specificity claims.

What to watch out for:

• Transfer across applications: An antibody’s clonality does not guarantee performance in every application—optimize for your method.
• Polyclonal lot variation: Revalidate when switching lots if your experiment depends on quantitative consistency.

What it means: Application(s) lists methods the supplier indicates this antibody has been used for. The value shown here is WB, IHC, ICC/IF, IP, FCM, FACS.

How to interpret here: Use WB, IHC, ICC/IF, IP, FCM, FACS as the closest available “validation scope” for your workflow. Performance is still dependent on sample type, prep conditions, and antigen abundance. Target is HSP25/HSP27—plan positive/negative controls that match expected expression and localization. Reactivity is Human, Mouse, Rat, Bovine, Dog, Guinea Pig, Hamster, Sheep—validation may not transfer across species without testing. Clonality is Monoclonal; this can influence robustness across fixation/denaturation conditions. Conjugate options include Unconjugated; ATTO 390; ATTO 488; ATTO 594; APC; Biotin; FITC; HRP; PCP; RPE; direct conjugates can simplify IF/FC workflows, while unconjugated formats provide flexibility. Supplier notes: Detects ~25kDa or ~27kDa. Recognizes HSP25 and HSP27, cross reacts with alpha B crystallin.—use these to interpret expected band sizes or specificity caveats in your readout.

Why it matters for buying:

• Risk reduction: Choosing an antibody already used in your application lowers the chance of extensive troubleshooting.
• Protocol compatibility: Some methods (IHC/IF/FC) are more sensitive to background than WB—validation matters more.
• Budget efficiency: Selecting the right format up-front (unconjugated vs conjugated) avoids re-purchasing.

Practical guidance:

• WB: Confirm expected molecular weight/isoforms; interpret multi-band patterns with controls (KO/KD when feasible).
• IHC: Fixation and antigen retrieval strongly affect staining; include isotype and secondary-only controls.
• IF/ICC: Optimize fixation/permeabilization; confirm localization with compartment markers when relevant.
• Flow cytometry: Verify whether the target is surface vs intracellular; plan compensation and FMO controls for gating.
• IP: Binding can be buffer-dependent; use lysis conditions that preserve epitope/complex and confirm pull-down by WB or MS.
• Record conditions: Document dilution, buffer, and sample prep for reproducibility and future reorders.

What to watch out for:

• Application transfer risk: Validation in one method doesn’t guarantee success in another.
• Matrix effects: Tissue autofluorescence, Fc receptor binding, and endogenous Ig can elevate background—controls matter.
• Lot changes: Revalidate after switching lots for critical quantitative comparisons.

What it means: Host is the species the antibody was raised in. The value shown here is Mouse.

How to interpret here: Use Mouse to select compatible secondary antibodies/detection reagents and to plan background controls. Use anti-mouse secondary reagents (and consider mouse-on-mouse background in mouse tissues). Isotype is IgG1 Kappa—match subclass-specific secondaries and isotype controls when needed. Conjugate options include Unconjugated; ATTO 390; ATTO 488; ATTO 594; APC; Biotin; FITC; HRP; PCP; RPE; direct conjugates may not require a secondary antibody. Reactivity is Human, Mouse, Rat, Bovine, Dog, Guinea Pig, Hamster, Sheep—remember: host = antibody origin; reactivity = species where the antigen is expected to be recognized. Applications listed: WB, IHC, ICC/IF, IP, FCM, FACS. Host choice matters most in IF/IHC/FC where secondary-driven background can dominate.

Why it matters for buying:

• Secondary compatibility: Host determines which secondaries and detection systems you can use.
• Multiplexing: Using primaries from different hosts can simplify multi-target staining.
• Background control: Certain sample types can bind secondaries non-specifically; host choice affects how hard background is to manage.

Practical guidance:

• Use cross-adsorbed secondaries: Helps reduce cross-reactivity in complex tissues or multiplex panels.
• Include secondary-only controls: Essential for interpreting IF/IHC background.
• Block Fc receptors when relevant: Particularly in immune-rich tissues or flow cytometry.

What to watch out for:

• Mouse-on-mouse background: Mouse primaries on mouse tissues can increase background—specialized strategies may be required.
• Subclass mismatches: Subclass-specific secondaries require a matching isotype/subclass.
• Cross-reactive secondaries: Poorly adsorbed secondaries can bind other primaries—validate multiplex reagents.