TXNDC17 Antibody

Basic Research Questions

• What is TXNDC17 and what are the key specifications for antibodies targeting this protein?

  TXNDC17 (Thioredoxin domain-containing protein 17), also known as TRP14 or TXNL5, is a 123-amino acid protein with a WCPDC motif that belongs to the thioredoxin family . This widely expressed protein has a thioredoxin fold and a TRX-like active-site sequence .

  Most commercial TXNDC17 antibodies share these key specifications:

  Characteristic	Typical Specifications
  Observed Molecular Weight	14 kDa
  Common Host	Rabbit
  Antibody Class	Polyclonal
  Reactivity	Human, Mouse, Rat
  Storage Conditions	-20°C (long-term), 4°C (short-term after reconstitution)
  Buffer Composition	PBS with 0.02% sodium azide and glycerol (typically 40-50%)

• What are the recommended applications and dilutions for TXNDC17 antibodies?

  TXNDC17 antibodies have been validated for multiple applications with specific recommended dilutions:

  Application	Recommended Dilution	Positive Detection Examples
  Western Blot (WB)	1:1000-1:50000	A549, HEK-293, HeLa, Jurkat, K-562, L02 cells; human kidney/uterus; mouse pancreas tissue
  Immunoprecipitation (IP)	0.5-4.0 μg per 1.0-3.0 mg protein lysate	HEK-293 cells
  Immunofluorescence (IF/ICC)	1:50-1:500	HepG2 cells, HeLa cells
  ELISA	1:5000-1:10000	Various human, mouse, rat samples
  Flow Cytometry	Typically 1 μg per 1×10^6 cells	JK cells
  Immunohistochemistry (IHC)	1:200-1:500	Human tissue sections

  It is recommended to optimize these dilutions for each specific experimental system to obtain optimal results .

• What is the optimal protocol for Western blotting using TXNDC17 antibodies?

  For optimal Western blot detection of TXNDC17:

  1. Sample Preparation:

     • Extract proteins from cells/tissues using RIPA buffer

     • Determine protein concentration using BCA Protein Assay Kit

     • Denature 20 μg protein at 100°C for 10 minutes in loading buffer

  2. Gel Electrophoresis & Transfer:

     • Separate proteins via SDS-PAGE (appropriate for 14 kDa detection)

     • Transfer to PVDF membrane

     • Block with skim milk powder for 2 hours at room temperature

  3. Antibody Incubation:

     • Primary antibody: Dilute TXNDC17 antibody 1:5000-1:50000 in blocking solution

     • Incubate overnight at 4°C

     • Wash membrane with TBS-T three times (15 minutes each)

     • Secondary antibody: HRP-labeled anti-rabbit IgG (typically 1:5000)

     • Incubate for 1 hour at room temperature

  4. Detection:

     • Wash extensively with TBS-T

     • Visualize using enhanced chemiluminescence reagent

     • Expected molecular weight: 14 kDa

• How should researchers validate TXNDC17 antibody specificity?

  Proper validation of TXNDC17 antibodies should include:

  1. Positive Controls:

     • Use cells with known TXNDC17 expression (A549, HEK-293, HeLa, Jurkat, K-562)

     • Include human kidney or uterus tissue, or mouse pancreas tissue lysates

  2. Negative Controls:

     • Include isotype control antibody (e.g., rabbit IgG at equivalent concentration)

     • Test with TXNDC17 knockdown cells (via shRNA)

  3. Cross-Reactivity Assessment:

     • Verify molecular weight (14 kDa) matches expected size

     • Some antibodies have been verified on protein arrays containing target protein plus 383 other non-specific proteins

  4. Multiple Detection Methods:

     • Confirm expression using at least two techniques (e.g., WB and IF/ICC)

     • Compare results between polyclonal antibodies from different sources