Filaggrin Recombinant Rabbit Monoclonal Antibody [PSH23-66]
cat.: HA751907
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, IF-Tissue, FC
Clonality: Monoclonal
Clone number: PSH23-66
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 435 kDa
Isotype: IgG
Immunogen: Recombinant protein within human Filaggrin aa 51-300.
Positive control: HeLa (Human cervical adenocarcinoma cells) cell lysate, A431 (Human epidermoid carcinoma skin squamous cells) cell lysate, SK-BR-3 (Human breast cancer cells) cell lysate, SK-MEL-28 (Human malignant melanoma cells) cell lysate, NCI-H441 (Human lung adenocarcinoma cells) cell lysate, NCI-H358 (Human lung adenocarcinoma cells) cell lysate, Mouse skin tissue lysate, Mouse pancreas tissue lysate, Mouse stomach tissue lysate, Rat brain tissue lysate, Rat stomach tissue lysate.
Subcellular location: Cytoplasmic granule.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  IF-Tissue
  FC
1:2,000-1:5,000
1:50
1:1000
1:200
1:200
Uniprot #: SwissProt: P20930 Human | P11088 Mouse
Entrez Gene: 24641 Rat
Alternative names: Filaggrin FLG
Images
HA751907_1.jpg Fig1: Western blot analysis of Filaggrin on different lysates with Rabbit anti-Filaggrin antibody (HA751907) at 1/2,000 dilution.

Lane 1: HeLa (Human cervical adenocarcinoma cells) cell lysate
Lane 2: A431 (Human epidermoid carcinoma skin squamous cells) cell lysate
Lane 3: SK-BR-3 (Human breast cancer cells) cell lysate
Lane 4: SK-MEL-28 (Human malignant melanoma cells) cell lysate
Lane 5: NCI-H441 (Human lung adenocarcinoma cells) cell lysate
Lane 6: NCI-H358 (Human lung adenocarcinoma cells) cell lysate

Lysates/proteins at 15 µg/Lane.
Exposure time: 3 minutes; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA751907, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 435 kDa
Observed band size: 37,77 kDa
HA751907_2.jpg Fig2: Western blot analysis of Filaggrin on different lysates with Rabbit anti-Filaggrin antibody (HA751907) at 1/5,000 dilution.

Lane 1: Mouse skin tissue lysate
Lane 2: Mouse pancreas tissue lysate
Lane 3: Mouse stomach tissue lysate
Lane 4: Rat brain tissue lysate
Lane 5: Rat stomach tissue lysate

Lysates/proteins at 30 µg/Lane.
Exposure time: 3 minutes; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA751907, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 435 kDa
Observed band size: 77 kDa
HA751907_3.jpg Fig3: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.

Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA751907, 1/50, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.

Counterstain: Beta tubulin (HA601187, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).
HA751907_4.jpg Fig4: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Skin
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751907, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA751907_5.jpg Fig5: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Liver
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751907, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.

Negative expression of Filaggrin protein in liver is consistent with the predicted expression pattern.
HA751907_6.jpg Fig6: Application: Immunofluorescence (IF-tissue)

Species: Human
Tissue: Skin
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× PBS
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751907, 1/200, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA751907_7.jpg Fig7: Application: Flow Cytometry (Intra)

Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.

Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA751907 (1/200) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.