Goat Anti-Human IgM/Bio
Ordering
Available Variants| Availability & Stock | Pack | SKU | Price (EUR) |
|---|---|---|---|
|
Available – Dispatch in 14 days
Global Network - International delivery
|
100ul | CAB.16427-100ul | €75.00 |
| Catalog No | CAB.16427 |
|---|---|
| Product Name | Goat Anti-Human IgM/Bio |
| Isotype | N/A |
| Calculated MW | 900kDa |
| Immunogen | N/A |
| Public Immunogen Range | N/A |
| Host | Goat |
| Clone Type | N/A |
| Reactivity | Human IgM |
| Application | IHC-F;FCM;IF;IHC-P;ELISA;WB |
| Subcellular Location | N/A |
| Purification Method | CABfinity purified by Protein G |
| Storage Buffer | N/A |
| Storage | Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. |
Background
IgM normally constitutes about 10% of serum immunoglobulins. IgM antibody is prominent in early immune responses to most antigens and is largely confined to plasma due to it’s large size. Monomeric IgM is expressed as a membrane bound antibody on the surface of B cells and as a pentamer when secreted by plasma cells. Due to it’s high valency IgM is more efficient than other isotypes is binding antigens with repeating epitopes (virus particles and red blood cells) and is more efficient than IgG in activiating the complement pathway. The gene for the mu constant region contains four domains separated by short intervening sequences.
Protocol / Instructions
**IHC-F Protocol** 1. Use fresh frozen sections fixed briefly with cold acetone or paraformaldehyde according to antigen sensitivity. 2. Bring slides to room temperature, wash in PBS, and block nonspecific binding before primary antibody incubation. 3. Apply Goat Anti-Human IgM/Bio at the optimized dilution for 1 hour at room temperature or overnight at 4°C. Recommended dilution: WB=1:1000-10000; ELISA=1:1000-10000; IHC-P=1:100-500; IHC-F=1:100-1000; IF=1:100-1000; FCM=1:100-1000. 4. Wash in PBS or TBS, apply enzyme- or fluorophore-conjugated secondary antibody, and protect from light when fluorescence detection is used. 5. Mount with compatible medium and evaluate signal localization against isotype or no-primary control sections. **FCM Protocol** Use assay conditions validated for FCM, apply Goat Anti-Human IgM/Bio at the recommended working dilution, include proper positive and negative controls, perform thorough wash steps, and document incubation time, temperature, and detection conditions for reproducible performance. Recommended dilution: WB=1:1000-10000; ELISA=1:1000-10000; IHC-P=1:100-500; IHC-F=1:100-1000; IF=1:100-1000; FCM=1:100-1000. **IF Protocol** Use assay conditions validated for IF, apply Goat Anti-Human IgM/Bio at the recommended working dilution, include proper positive and negative controls, perform thorough wash steps, and document incubation time, temperature, and detection conditions for reproducible performance. Recommended dilution: WB=1:1000-10000; ELISA=1:1000-10000; IHC-P=1:100-500; IHC-F=1:100-1000; IF=1:100-1000; FCM=1:100-1000. **IHC-P Protocol** 1. Use formalin-fixed, paraffin-embedded sections mounted on charged slides. Bake, deparaffinize in xylene, and rehydrate through graded ethanol to water. 2. Perform heat-induced epitope retrieval in citrate or EDTA buffer, then cool and rinse in PBS or TBS. 3. Quench endogenous peroxidase if chromogenic detection will be used, and block nonspecific binding with serum or protein block for 20–30 minutes. 4. Incubate sections with Goat Anti-Human IgM/Bio in antibody diluent for 1 hour at room temperature or overnight at 4°C. Recommended dilution: WB=1:1000-10000; ELISA=1:1000-10000; IHC-P=1:100-500; IHC-F=1:100-1000; IF=1:100-1000; FCM=1:100-1000. 5. Wash thoroughly, apply the appropriate secondary detection system, develop with DAB or equivalent substrate, counterstain with hematoxylin, dehydrate, mount, and compare specific staining with negative control tissue. **ELISA Protocol** 1. Coat the plate with antigen or capture reagent according to assay design, block with protein-containing buffer to reduce nonspecific binding, and incubate with Goat Anti-Human IgM/Bio at the optimized working dilution. Recommended dilution: WB=1:1000-10000; ELISA=1:1000-10000; IHC-P=1:100-500; IHC-F=1:100-1000; IF=1:100-1000; FCM=1:100-1000. 2. Wash between each incubation using PBS or TBS containing a low concentration of Tween-20. 3. Add enzyme-conjugated secondary antibody, develop with appropriate substrate, stop the reaction at the recommended endpoint, and read absorbance promptly. 4. Generate a control-based interpretation using blank, negative, and positive wells. **Western Blotting Protocol** 1. Prepare lysates in SDS-compatible lysis buffer containing protease inhibitors, clarify by centrifugation, and denature samples at 95–100°C for 5 minutes. 2. Resolve proteins by SDS-PAGE and transfer to PVDF or nitrocellulose membrane. 3. Block with 5% BSA or nonfat milk in TBST for 1 hour at room temperature. 4. Incubate the membrane with Goat Anti-Human IgM/Bio diluted in antibody dilution buffer overnight at 4°C with gentle agitation. Recommended dilution: WB=1:1000-10000; ELISA=1:1000-10000; IHC-P=1:100-500; IHC-F=1:100-1000; IF=1:100-1000; FCM=1:100-1000. 5. Wash 3 times in TBST, then incubate with species-appropriate HRP-conjugated secondary antibody for 1 hour at room temperature. 6. Wash again, develop with ECL.
Precautions
For research use only. Store under the recommended conditions and avoid repeated freeze-thaw cycles. Mix gently before use, centrifuge briefly if needed, and validate dilution, retrieval, fixation, permeabilization, and incubation conditions in your assay system. Use appropriate positive, negative, and no-primary controls, and handle all biological samples using standard laboratory biosafety procedures.
Validation Images