Antigen Retrieval Buffer (100X Citrate Buffer)(AB93678)

Antigen Retrieval Buffer (100X Citrate Buffer) is a special solution that enables rehydration and antigen retrieval to be performed in formalin-fixed, paraffin-embedded tissue sections with one step. It is optimal for use with primary antibodies that require citrate buffer pretreatment.

This product contains detergent for emulsification of the paraffin.

1X Dilution:

The 100X stock solution should be diluted 100-fold with distilled water before use.

Protocol:

1. Place paraffin-embedded slides in 1x Antigen Retrieval Buffer; cover with a vented plastic wrap, place in microwave and set high power to boil and then set low power to keep it boiling for 10 min. Let the sections cool in the microwave for at least 20 min.

2. Wash sections with hot tap water for 1 min.

3. Wash sections in buffer for 2 x 3 minutes.

4. Continue with IHC protocol as normal.

Other kits and reagents for IHC

Other antigen retrieval buffers include: Tris-EDTA buffer pH 9.0 ab93684, EDTA buffer pH 8.0 ab93680, Tris buffer pH 10.0 ab93682, or see the full list of antigen retrieval buffers and enzymes.

Find more kits and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in the IHC kits and reagents guide.

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Fibrillarin with ab314506 at 1/2000 (0.249 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Nucleolus staining on human tonsil The section was incubated with ab314506 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0)

Immunohistochemistry analysis of paraffin-embedded human cerebellum tissue sections labelling GFAP with ab4648 at 1/500 dilution (1 mg/mL). Sections were stained with ab4648 using the HRP/DAB staining. Sections were counterstained with hematoxylin/eosin. Antigen retrieval was heat mediated using Antigen retrieval buffer (100X citrate buffer) (pH 6.0) (ab93678) for 15 minutes. To the right is a region of cerebellar molecular layer containing the prominent cytoskeletal fibers of Bergmann glia which are strongly positive for GFAP. The middle shows a region of the granular layer and to the left is white matter, both of which contain GFAP positive astrocytes.The immunostaining was performed with the Vector ImmPress rat adsorbed horse anti-mouse IgG detection kit.

Immunohistochemical analysis of paraffin-embedded (A) FaDu (human pharyngeal carcinoma epithelial cell) cell pellet, (B) A431 (human epidermoid carcinoma epithelial cell) cell pellet and (C) K-562 (human chronic myelogenous leukemia lymphoblast) cell pellet labeling CELSR1 with ab322043 at 1/50 (10.36 μg/ml) dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). 

Positive staining on (A) FaDu cell pellet and (B) A431 cell pellet, no staining on (C) K-562 cell pellet. 

The section was incubated with ab322043 at 4°C overnight. 

Counterstained with Hematoxylin. 

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP). 

Heat mediated antigen retrieval was performed with ab93678 (citrate buffer, pH 6.0)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Fibrillarin with ab314506 at 1/2000 (0.249 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Nucleolus staining on mouse liver. The section was incubated with ab314506 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Fibrillarin with ab314506 at 1/2000 (0.249 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Nucleolus staining on rat liver. The section was incubated with ab314506 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Fibrillarin with ab314506 at 1/2000 (0.249 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Nucleolus staining on mouse testis. The section was incubated with ab314506 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0)

ab93678 Antigen Retrieval Bufer 100X Citrate Buffer

Nuclear OPN expression in explanted hearts but not endomyocardial biopsies (EMBs)In Patient #1, nuclear OPN (shown in white) in ACTA1-positive cardiomyocytes (shown in red) is visible in failing native and transplanted hearts, but in none of the three serial follow-up EMB samples collected at 1 week, 1 month, and 1 year post first transplant. Representative confocal z-stack images are shown. Scale bar = 20 μm.

Immunohistochemical staining of GFP in the indicated genotypes (5 mice per group) : Immunohistochemical staining of GFP in the indicated genotypes (n=5 mice per group) : The paraffin-embedded sections were deparaffinized with xylene and alcohol and subjected to antigen retrieval by citrate buffer (Abcam, Cat# ab93678), Tris-EDTA buffer (Abcam, Cat# ab93684), EDTA buffer (Abcam, Cat# ab93680), or Tris buffer (Abcam, Cat# ab93682) in a 100°C cooker for 30 min. After blocking, the sections were incubated with primary antibodies at 4°C overnight. 1 : 500, Chicken anti-GFP, Abcam, Cat#ab13970. the mouse tissue sections were developed with an IHC Detection kit (Thermo Fisher, Cat#3600) according to the manufacturer's instructions. 

Image from Wang et al., Nat Commun., 11(1):220. doi: 10.1038/s41467-019-14046-9. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/