RBM12B Mouse mAb: Protocol & Usage Guide for Research

RBM12B Mouse mAb is a monoclonal antibody targeting RNA Binding Motif Protein 12B, supplied in PBS formulation and intended for use in standard immunological research applications.

Product SKU

Ab350679

Target Protein

RBM12B (RNA Binding Motif Protein 12B)

Host Species

Mouse

Antibody Format

Monoclonal Antibody (mAb)

Formulation

PBS Only

Source Reference

absea#KC-41672

Overview of RBM12B and Antibody Specifications

RBM12B (RNA Binding Motif Protein 12B) is a member of the RNA-binding protein family characterized by the presence of RNA recognition motif (RRM) domains. RNA-binding proteins play essential roles in post-transcriptional gene regulation, including pre-mRNA splicing, mRNA stability, transport, and translation. RBM12B is expressed in various mammalian tissues and has been studied in the context of cellular development and differentiation.

The RBM12B Mouse mAb (Catalog: Ab350679) is a monoclonal antibody raised in mouse against the RBM12B protein. It is supplied in PBS (phosphate-buffered saline) without additional stabilizers or preservatives, making it suitable for a variety of downstream immunological assays. The source reference for this product is absea#KC-41672.

Storage, Handling, and Reconstitution

Storage Conditions: Upon receipt, store the RBM12B Mouse mAb at –20 °C for long-term storage. For short-term use (up to 1 month), the antibody may be kept at 4 °C. Repeated freeze-thaw cycles should be avoided to maintain antibody integrity; aliquoting the stock solution upon first use is strongly recommended.

Handling: As the product is formulated in PBS only, users should take care to avoid microbial contamination. Work under sterile conditions where possible, and use low-protein-binding tubes for aliquots. Do not dilute the antibody until immediately prior to use.

Reconstitution: This product is supplied in liquid form in PBS; no reconstitution is required. If the product has been lyophilized (refer to the specific vial label), reconstitute with the volume of sterile distilled water or PBS indicated on the datasheet and allow to stand for 10 minutes at room temperature before use.

Recommended Protocols for Key Applications

Western Blot (WB):

1. Prepare total cell or tissue lysate using standard RIPA buffer supplemented with protease inhibitors.
2. Separate proteins by SDS-PAGE and transfer to a PVDF or nitrocellulose membrane.
3. Block the membrane with 5% non-fat dry milk or BSA in TBST for 1 hour at room temperature.
4. Incubate with RBM12B Mouse mAb at the recommended dilution (refer to product datasheet; typical starting dilution 1:500–1:2000) overnight at 4 °C.
5. Wash 3× with TBST (10 minutes each), then incubate with an appropriate HRP-conjugated secondary anti-mouse antibody for 1 hour at room temperature.
6. Detect signal using an ECL substrate system and image accordingly.

Immunohistochemistry (IHC):

1. Fix tissue sections in 10% neutral-buffered formalin and embed in paraffin (FFPE) or use frozen sections.
2. Perform antigen retrieval using citrate buffer (pH 6.0) or EDTA buffer (pH 9.0) as appropriate.
3. Block endogenous peroxidase with 3% H₂O₂ and block non-specific binding with 5–10% normal serum.
4. Incubate with RBM12B Mouse mAb at the recommended dilution overnight at 4 °C in a humidified chamber.
5. Apply secondary antibody and detection reagent per your chosen kit protocol, counterstain with hematoxylin, and mount.

Immunofluorescence (IF):

1. Fix cells on coverslips with 4% paraformaldehyde for 15 minutes, then permeabilize with 0.1–0.3% Triton X-100 in PBS.
2. Block with 5% BSA or normal serum for 30–60 minutes.
3. Incubate with RBM12B Mouse mAb at recommended dilution for 1 hour at room temperature or overnight at 4 °C.
4. Wash 3× with PBS, then incubate with a fluorescently labeled anti-mouse secondary antibody for 1 hour at room temperature, protected from light.
5. Mount with DAPI-containing mounting medium and image by fluorescence microscopy.

Troubleshooting Common Issues

No Signal or Weak Signal: Verify antibody storage conditions and check for repeated freeze-thaw damage. Increase primary antibody concentration or extend incubation time. Confirm that the target protein (RBM12B) is expressed in the cell line or tissue tested. Optimize antigen retrieval conditions for IHC applications.

High Background: Increase blocking time or optimize blocking agent concentration. Increase washing stringency (more washes, longer wash times, or addition of 0.05–0.1% Tween-20 to wash buffer). Reduce primary antibody concentration. For IHC, ensure endogenous peroxidase is fully quenched.

Non-Specific Bands (WB): Confirm correct gel percentage for expected molecular weight of RBM12B. Reduce primary antibody concentration. Include a negative control (lysate from RBM12B-knockout or non-expressing cells). Check for cross-reactivity with closely related RBM family members.

Inconsistent Results: Ensure all reagents are equilibrated to room temperature before use. Standardize protein loading using a housekeeping protein control. Confirm lot-to-lot consistency of secondary antibody and detection reagents.

Frequently asked questions

What formulation is the RBM12B Mouse mAb supplied in?

The RBM12B Mouse mAb (Ab350679) is supplied in PBS (phosphate-buffered saline) only, without added stabilizers or preservatives. Users should aliquot upon first use and avoid repeated freeze-thaw cycles.

What are the recommended storage conditions for Ab350679?

For long-term storage, keep the antibody at –20 °C. For short-term use within one month, storage at 4 °C is acceptable. Always avoid repeated freeze-thaw cycles by preparing single-use aliquots.

Which applications is the RBM12B Mouse mAb suitable for?

Based on general monoclonal antibody use, the RBM12B Mouse mAb may be applicable for Western Blot (WB), Immunohistochemistry (IHC), and Immunofluorescence (IF). Users should consult the specific product datasheet for validated applications and recommended dilutions, as confirmed application data is subject to review.

What is the source reference for this antibody product?

The RBM12B Mouse mAb (Ab350679) is referenced under absea#KC-41672. Please cite this reference number when contacting Aladdin Scientific technical support for product-specific queries.

How should I troubleshoot if I observe high background in Western Blot?

High background in WB typically results from insufficient blocking or excessive primary antibody concentration. Increase blocking time, optimize the blocking agent, reduce primary antibody concentration, and increase washing stringency by adding 0.05–0.1% Tween-20 to the wash buffer.

References

1. Product datasheet: RBM12B Mouse mAb, Ab350679, absea#KC-41672, Aladdin Scientific.
2. Lunde BM, Moore C, Varani G. RNA-binding proteins: modular design for efficient function. Nat Rev Mol Cell Biol. 2007;8(6):479-490. doi:10.1038/nrm2178

Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.