Iba1 Recombinant Antibody [JM36-62] - Chicken IgY (Chimeric)

Application: IHC-Fr

Species: Mouse

Site: Cerebellum

Sample: Frozen section

Antibody concentration: 1/500 (Iba1, HA601376, green); 1/500 (MAG, HA721818, red)

Antigen retrieval: Not required

Application: IHC-Fr

Species: Mouse

Site: Cerebellum

Sample: Frozen section

Antibody concentration: 1/500 (Iba1, HA601376, green); 1/500 (PSD95, ET1602-20, red)

Antigen retrieval: Not required

Application: IHC-Fr

Species: Mouse

Site: Spleen

Sample: Frozen section

Antibody concentration: 1/500 (Iba1, HA601376, green); 1/1,000 (F4/80, HA721520, red)

Antigen retrieval: Not required

Application: IHC-Fr

Species: Rat

Site: Spleen

Sample: Frozen section

Antibody concentration: 1/500 (Iba1, HA601376, green); 1/1,000 (F4/80, HA721520, red)

Antigen retrieval: Not required

☑ Relative expression (RE)

Western blot analysis of Iba1 on different lysates with Chicken anti-Iba1 antibody (HA601376) at 1/5,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: HEK-293 cell lysate (negative)
Lane 3: Mouse spleen tissue lysate
Lane 4: Rat spleen tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 17 kDa
Observed band size: 17 kDa

Exposure time: 3 minutes; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601376) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat anti-Chicken IgY - HRP Secondary Antibody (HA1146) at 1/10,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of C6 cells labeling Iba1 with Chicken anti-Iba1 antibody (HA601376) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Chicken anti-Iba1 antibody (HA601376) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Chicken IgY (iFluor™ 647, HA1142) was used as the secondary antibody at 1/250 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Flow cytometric analysis of THP-1 cells labeling Iba1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA601376, 1/1,000) (red). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 750 conjugate-Goat anti-Chicken IgY Secondary antibody (HA1145) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).